Heterocyclic Compounds and Uses Thereof

ABSTRACT

Heterocyclic entities that modulate PI3 kinase activity, pharmaceutical compositions containing the heterocyclic entities, and methods of using these chemical entities for treating diseases and conditions associated with PI3 kinase activity are described herein.

This application claims the benefit of U.S. Provisional Application Ser.No. 61/234,617, filed Aug. 17, 2009, which is hereby incorporated byreference in its entirety.

BACKGROUND OF THE INVENTION

The activity of cells can be regulated by external signals thatstimulate or inhibit intracellular events. The process by whichstimulatory or inhibitory signals are transmitted into and within a cellto elicit an intracellular response is referred to as signaltransduction. Over the past decades, cascades of signal transductionevents have been clucidated and found to play a central role in avariety of biological responses. Defects in various components of signaltransduction pathways have been found to account for a vast number ofdiseases, including numerous forms of cancer, inflammatory disorders,metabolic disorders, vascular and neuronal diseases (Gaestel et al.Current Medicinal Chemistry (2007) 14:2214-2234).

Kinases represent a class of important signaling molecules. Kinases cangenerally be classified into protein kinases and lipid kinases, andcertain kinases exhibit dual specificities. Protein kinases are enzymesthat phosphorylate other proteins and/or themselves (i.e.,autophosphorylation). Protein kinases can be generally classified intothree major groups based upon their substrate utilization: tyrosinekinases which predominantly phosphorylate substrates on tyrosineresidues (e.g., erb2, PDGF receptor, EGF receptor, VEGF receptor, src,abl), serine/threonine kinases which predominantly phosphorylatesubstrates on serine and/or threonine residues (e.g., mTorC1, mTorC2,ATM, ATR, DNA-PK, Akt), and dual-specificity kinases which phosphorylatesubstrates on tyrosine, serine and/or threonine residues.

Lipid kinases are enzymes that catalyze the phosphorylation of lipidswithin cells. These enzymes, and the resulting phosphorylated lipids andlipid-derived biologically active organic molecules, play a role in manydifferent physiological processes, including cell proliferation,migration, adhesion, and differentiation. A particular group of lipidkinases comprises membrane lipid kinases, i.e., kinases that catalyzethe phosphorylation of lipids contained in or associated with cellmembranes. Examples of such enzymes include phosphinositide(s) kinases(such as PI3-kinases, PI4-Kinases), diacylglycerol kinases, andsphingosine kinases.

The phosphoinositide 3-kinases (PI3Ks) signaling pathway is one of themost highly mutated systems in human cancers. PI3K signaling is involvedin many other disease states including allergic contact dermatitis,rheumatoid arthritis, osteoarthritis, inflammatory bowel diseases,chronic obstructive pulmonary disorder, psoriasis, multiple sclerosis,asthma, disorders related to diabetic complications, and inflammatorycomplications of the cardiovascular system such as acute coronarysyndrome.

PI3Ks are members of a unique and conserved family of intracellularlipid kinases that phosphorylate the 3′-OH group onphosphatidylinositols or phosphoinositides. The PI3K family comprises 15kinases with distinct substrate specificities, expression patterns, andmodes of regulation (Katso et al., 2001). The class I PI3Ks (p110α,p110β, p110δ, and p110γ) are typically activated by tyrosine kinases orG-protein coupled receptors to generate PIP3, which engages downstreameffectors such as those in the pathways of Akt/PDK1, mTOR, the Tecfamily kinases, and the Rho family GTPases. The class II and III PI3-Ksplay a key role in intracellular trafficking through the synthesis ofPI(3)P and PI(3,4)P₂.

The alpha (α) isoform of PI3K has been implicated, for example, in avariety of human cancers. Angiogenesis has been shown to selectivelyrequire the a isoform of PI3K in the control of endothelial cellmigration. (Graupera et al, Nature 2008; 453; 662-6). Mutations in thegene coding for PI3K α or mutations which lead to upregulation of PI3K αare believed to occur in many human cancers such as lung, stomach,endometrial, ovarian, bladder, breast, colon, brain and skin cancers.Often, mutations in the gene coding for PI3K α are point mutationsclustered within several hotspots in helical and kinase domains, such asE542K, E545K, and H1047R. Many of these mutations have been shown to beoncogenic gain-of-function mutations. Because of the high rate of PI3K αmutations, targeting of this pathway may provide valuable therapeuticopportunities. While other PI3K isoforms such as PI3K δ or PI3K γ areexpressed primarily in hematopoietic cells, PI3K α, along with PI3K β,is expressed constitutively.

The delta (δ) isoform of class I PI3K has been implicated, inparticular, in a number of diseases and biological processes. PI3K δ isexpressed primarily in hematopoietic cells including leukocytes such asT-cells, dendritic cells, neutrophils, mast cells, B-cells, andmacrophages. PI3K δ is integrally involved in mammalian immune systemfunctions such as T-cell function, B-cell activation, mast cellactivation, dendritic cell function, and neutrophil activity. Due to itsintegral role in immune system function, PI3K δ is also involved in anumber of diseases related to undesirable immune response such asallergic reactions, inflammatory diseases, inflammation mediatedangiogenesis, rheumatoid arthritis, auto-immune diseases such as lupus,asthma, emphysema and other respiratory diseases. Other class I PI3Kinvolved in immune system function includes PI3K γ, which plays a rolein leukocyte signaling and has been implicated in inflammation,rheumatoid arthritis, and autoimmune diseases such as lupus.

Downstream mediators of the PI3K signal transduction pathway include Aktand mammalian target of rapamycin (mTOR). Akt possesses a pleckstrinhomology (PH) domain that binds PIP3, leading to Akt kinase activation.Akt phosphorylates many substrates and is a central downstream effectorof PI3K for diverse cellular responses. One important function of Akt isto augment the activity of mTOR, through phosphorylation of TSC2 andother mechanisms. mTOR is a serine-threonine kinase related to the lipidkinases of the PI3K family. mTOR has been implicated in a wide range ofbiological processes including cell growth, cell proliferation, cellmotility and survival. Disregulation of the mTOR pathway has beenreported in various types of cancer. mTOR is a multifunctional kinasethat integrates growth factor and nutrient signals to regulate proteintranslation, nutrient uptake, autophagy, and mitochondrial function.

Dysregulation of signaling pathways mediated by many other kinases is akey factor in the development of human diseases. Aberrant or excessiveprotein kinase activity or expression has been observed in many diseasestates including benign and malignant proliferative diseases, disorderssuch as allergic contact dermatitis, rheumatoid arthritis,osteoarthritis, inflammatory bowel diseases, chronic obstructivepulmonary disorder, psoriasis, multiple sclerosis, asthma, disordersrelated to diabetic complications, and inflammatory complications of thecardiovascular system such as acute coronary syndrome.

As such, kinases particularly lipid kinases such as PI3Ks and proteinkinases such as mTor are prime targets for drug development. The presentinvention addresses a need in the art by providing a new class of kinaseinhibitors.

SUMMARY OF THE INVENTION

In one embodiment, the invention provides a compound of Formula I:

or its pharmaceutically acceptable salts thereof, wherein

-   -   X is O or S or N;    -   W¹ is N, NR³, CR³, or C═O, W² is N, NR⁴, CR⁴, or C═O, W³ is N,        NR⁵ or CR⁵, W⁴ is N, C═O or CR⁶, wherein no more than two N        atoms and no more than two C═O groups are adjacent;    -   W⁵ is N or CR⁷;    -   W⁶ is N or CR⁸;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety,    -   or R³ and R⁴ taken together form a cyclic moiety; and    -   R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, alkyl,        heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,        aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,        heterocycloalkyloxy, amido, amino, acyl, acyloxy,        alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,        phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are taken        together with nitrogen to form a cyclic moiety.

In some embodiments, the compound has the formula:

In still other embodiments of compounds of Formula I, W¹ is CR³, W² isCR⁴, W³ is CR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸; W¹ is N, W² is CR⁴,W³ is CR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸; or W¹ is CR³, W² is N, W³is CR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸. In some embodiments, W⁵ andW⁶ are CH. In other embodiments, R² is amino. In yet other embodiments,R¹ is H.

In some embodiments of compounds of Formula I, W¹ is CR³ and R³ isalkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,heterocycloalkyloxy, amido, amino, acyl, acyloxy, alkoxycarbonyl,sulfonamido, halo, cyano, hydroxy, nitro, phosphate, urea, carbonate, orNR′R″ wherein R′ and R″ are taken together with nitrogen to form acyclic moiety. In some embodiments, R³ is aryl, heteroaryl,heterocycloalkyl, or NR′R″ wherein R′ and R″ are taken together withnitrogen to form a cyclic moiety. In other embodiments, W¹ is CR³ and W²is CR⁴; and R³ and R⁴ taken together form a cyclic moiety. In oneembodiment, R³ and R⁴ taken together form a 5-membered heterocyclicring. In some embodiments, W⁴ is N. In other embodiments, X is O.

Compounds of Formula I are also provided wherein the cyclic moiety isfurther substituted with alkyl, heteroalkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino, acyl,acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are takentogether with nitrogen to form a cyclic moiety.

In another aspect, the invention provides a compound of Formula II:

or its pharmaceutically acceptable salts thereof, wherein

-   -   X is O or S or N;    -   W¹ is S, N, NR³ or CR³, W² is N or CR⁴, W³ is S, N or CR⁵, W⁴ is        N or C, and W⁷ is N or C, wherein no more than two N atoms and        no more than two C═O groups are adjacent;    -   W⁵ is N or CR⁷;    -   W⁶ is N or CR⁸;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   or R³ and R⁴ taken together form a cyclic moiety; and    -   R⁵, R⁷ and R⁸ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

In one embodiment, the compound of the present invention has theformula:

In other embodiments, W¹ is CR³, W² is CR⁴, W³ is N, W⁴ is N, W⁵ is CR⁷,and W⁶ is CR⁸; W¹ is CR³, W² is CR⁴, W³ is N, W⁴ is N, W⁵ is CR⁷, and W⁶is CR⁸; W¹ is CR³, W² is CR⁴, W³ is N, W⁴ is N, W⁵ is N, and W⁶ is CR⁸;W¹ is NR³, W² is CR⁴, W³ is N, W⁴ is C, W⁵ is CR⁷, and W⁶ is CR⁸; W¹ isS, W² is CR⁴, W³ is N, W⁴ is C, W⁵ is CR⁷, and W⁶ is CR⁸; or W¹ is CR³,W² is CR⁴, W³ is S, W⁴ is C, W⁵ is N, and W⁶ is N. In some embodiments,W⁵ and W⁶ are CH. In other embodiments, R² is amino. In still otherembodiments, R¹ is H.

In some embodiments, W¹ is CR³ and R³ is alkyl, heteroalkyl, alkenyl,alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino, acyl,acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are takentogether with nitrogen to form a cyclic moiety. In other embodiments, R³is aryl, heteroaryl, heterocycloalkyl, or NR′R″ wherein R′ and R″ aretaken together with nitrogen to form a cyclic moiety. In someembodiments, W¹ is CR³ and W² is CR⁴, and R³ and R⁴ taken together forma cyclic moiety. In other embodiments, R³ and R⁴ taken together form a5-membered heterocyclic ring.

In some embodiments, W⁴ is N. In other embodiments, the cyclic moiety isfurther substituted with alkyl, heteroalkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino, acyl,acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are takentogether with nitrogen to form a cyclic moiety. In still otherembodiments, R¹ is alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,heterocycloalkyloxy, amido, amino, acyl, acyloxy, alkoxycarbonyl,sulfonamido, halo, cyano, hydroxy, nitro, phosphate, urea, carbonate, orNR′R″ wherein R′ and R″ are taken together with nitrogen to form acyclic moiety. In some embodiments of the compound of Formula II, X isO. In other embodiments of compounds of Formula II, W⁷ is C.

The invention further provides a compound of Formula III:

or its pharmaceutically acceptable salts thereof, wherein

-   -   W¹ is CR³, W² is C-benzoxazolyl substituted with R² and W³ is S;    -   W¹ is CR³, W² is C-benzoxazolyl substituted with R² and W³ is        CR⁵;    -   W¹ is N or NR³, W² is CR⁴, and W³ is C-benzoxazolyl substituted        with R²;    -   W¹ is CR³, W² is CR⁴, and W³ is C-benzoxazolyl substituted with        R²; or    -   W¹ is N or NR³, W² is NR⁴, and W³ is C-benzoxazolyl substituted        with R²;    -   X is N;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, alkyl,        heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,        aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,        heterocycloalkyloxy, amido, amino, acyl, acyloxy,        alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,        phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are taken        together with nitrogen to form a cyclic moiety.

In some embodiments of the compound of Formula III, the compound is:

wherein W¹ is CR³ or NR³ and W² is CR⁴.

In one embodiment, the compound is:

wherein W¹ is CR³ or NR³ and W³ is CR⁵.

In another aspect, the invention provides a compound of Formula IV:

or its pharmaceutically acceptable salts thereof, wherein:

-   -   W¹ is N, NR³, CR³, or C═O; W² is N, NR⁴, CR⁴, or C═O; W³ is N,        NR⁵ or CR⁵; W⁴ is N, C═O or CR⁶, wherein no more than two N        atoms and no more than two C═O groups are adjacent;    -   W⁵ is N or CR⁷;    -   W⁶ is N or CR⁸;    -   W^(a) and W^(b) are independently N or CR⁹;    -   one of W^(c) and W^(d) is N, and the other is O, NR¹⁰, or S;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.    -   or R³ and R⁴ taken together form a cyclic moiety;    -   R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, alkyl,        heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,        aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,        heterocycloalkyloxy, amido, amino, acyl, acyloxy,        alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,        phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are taken        together with nitrogen to form a cyclic moiety;    -   R⁹ is alkyl or halo; and    -   R¹⁰ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

In some embodiments of the compound of Formula IV, W¹ is CR³, W² is CR⁴,W³ is CR³, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸; W¹ is N, W² is CR⁴, W³ isCR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸; or W¹ is CR³, W² is N, W³ isCR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸. In some embodiments of thecompound of Formula IV, W^(b) is N. In other embodiments, W^(a) is CR⁹and R⁹ is alkyl.

The invention also provides a compound of Formula V:

or its pharmaceutically acceptable salts thereof, wherein

-   -   W¹ is S, N, NR³ or CR³, W² is N or CR⁴, W³ is S, N or CR⁵, W⁴ is        N or C, and W⁷ is N or C, wherein no more than two N atoms and        no more than two C═O groups are adjacent;    -   W⁵ is N or CR⁷;    -   W⁶ is N or CR⁸;    -   W^(a) and W^(b) are independently N or CR⁹;    -   one of W^(c) and W^(d) is N, and the other is O, NR¹⁰, or S;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   or R³ and R⁴ taken together form a cyclic moiety;    -   R⁵, R⁷ and R⁸ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R⁹ is alkyl or halo; and    -   R¹⁰ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

In some embodiments of the compound of Formula V, W¹ is CR³, W² is CR⁴,W³ is N, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸. In other embodiments, W¹ isCR³, W² is CR⁴, W³ is N, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸. In otherembodiments, W¹ is CR³, W² is CR⁴, W³ is N, W⁴ is N, W⁵ is N, and W⁶ isCR⁸. In still other embodiments, W¹ is NR³, W² is CR⁴, W³ is N, W⁴ is C,W⁵ is CR⁷, and W⁶ is CR⁸. In other embodiments, W¹ is S, W² is CR⁴, W³is N, W⁴ is C, W⁵ is CR⁷, and W⁶ is CR⁸. In other embodiments, W¹ isCR³, W² is CR⁴, W³ is S, W⁴ is C, W⁵ is N, and W⁶ is N.

In some embodiments of the compound of Formula V, W^(b) is N. In otherembodiments, W^(a) is CR⁹ and R⁹ is alkyl.

The invention further provides compounds of Formula V-A and V-B:

or its pharmaceutically acceptable salts thereof, wherein

-   -   W¹ is CR³;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   and R³ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

Also provided herein are compounds of Formula V-C and V-D:

or its pharmaceutically acceptable salts thereof, wherein

-   -   W¹ is CR³;    -   W⁵ is N or CR⁷;    -   W^(a) and W^(b) are independently N or CR⁹;    -   one of W^(c) and W^(d) is N, and the other is O, NR¹⁰, or S;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R⁷ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R⁹ is alkyl or halo; and    -   R¹⁰ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

In some embodiments of the compound of Formula V-C or V-D, W^(b) is N.In other embodiments, W^(a) is CR⁹ and R⁹ is alkyl.

Also provided herein is a compound of Formula VI:

or its pharmaceutically acceptable salts thereof, wherein

-   -   W¹ is CR³; W₂ is CR⁴;    -   W^(a) is CH or N;    -   R¹ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ is alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,        heterocycloalkyl, aryl, arylalkyl, heteroarylalkyl, alkoxy,        heterocycloalkyloxy, amido, amino, acyl, acyloxy,        alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,        phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are taken        together with nitrogen to form a cyclic moiety;    -   R⁴ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   or R³ and R⁴ taken together form a cyclic moiety; and    -   R¹⁰ and R¹¹ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

The invention further provides a compound of Formula VII:

-   -   or a pharmaceutically acceptable salt thereof, wherein    -   X₁ is CR³, NR³, or S;    -   X₂ is CR⁴, NR⁴, CR⁴        CR⁵, or CR⁴        NR⁵;    -   X₃ and X₄ are independently C or N;    -   X₅ is CR⁶, NR⁶, or S;    -   X₄ is CR⁷, NR⁷, CR⁷        CR⁸, or CR⁷        NR⁸;    -   W^(a) and W^(b) are independently N or CR⁹;    -   one of W^(c) and W^(d) is N, and the other is O, NR¹⁰, or S;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   or R³ and R⁴ taken together form a cyclic moiety;    -   R⁵, R⁶, R⁷, and R⁸ are independently hydrogen, alkyl,        heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,        aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,        heterocycloalkyloxy, amido, amino, acyl, acyloxy,        alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,        phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are taken        together with nitrogen to form a cyclic moiety;    -   R⁹ is alkyl or halo; and    -   R¹⁰ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

In some embodiments of the compound of Formula VII, W^(b) is N. In otherembodiments, W^(a) is CR⁹ and R⁹ is alkyl.

In yet another embodiment, the present invention provides a compositioncomprising a pharmaceutically acceptable excipient and one or morecompound disclosed herein. In some embodiments, the composition is aliquid, solid, semi-solid, gel, or an aerosol form.

In still yet another embodiment, the present invention provides a methodfor inhibiting a phosphatidyl inositol-3 kinase (PI3 kinase),comprising: contacting the PI3 kinase with an effective amount of acompound disclosed herein. In some embodiments, the PI3 kinase is PI3kinase alpha. The step of contacting may further comprise contacting acell that expresses one or more type I PI3 kinases, including PI3kinease alpha. In some embodiments, the method further comprisesadministering a second therapeutic agent to the cell.

The present invention also provides a method for treating a conditionassociated with PI3 kinase, comprising administering to a subject inneed thereof an effective amount of the compound disclosed herein. Insome embodiments, the condition associated with PI3 kinase is selectedfrom the group consisting of asthma, emphysema, bronchitis, psoriasis,allergy, anaphylaxis, rheumatoid arthritis, graft versus host disease,lupus erythematosus, psoriasis, restenosis, benign prostatichypertrophy, diabetes, pancreatitis, proliferative glomerulonephritis,diabetes-induced renal disease, inflammatory bowel disease,atherosclerosis, eczema, scleroderma, diabetes, diabetic retinopathy,retinopathy of prematurity, age-related macular degeneration,hemangioma, glioma, melanoma, Kaposi's sarcoma and ovarian, breast,lung, pancreatic, prostate, colon and epidermoid cancer.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1 illustrates the selectivity and potency of inhibition of anexemplary compound of the invention (e.g., compound 54) with respect toa number of kinases.

FIG. 2 shows that an exemplary compound of the invention (e.g., compound54) inhibits the PI3K pathway and the proliferation of tumor cells witha PI3Kα mutation.

FIG. 3 shows the inhibition of angiogenesis by a compound of theinvention, and comparison to the activity of a non-specific inhibitor ofPI3Ks (Pan-PI3K Inh).

FIG. 4 shows the in vivo efficacy of a compound of the invention bycomparison to a Pan-PI3K inhibitor, as measured by tumor weight anddownstream markers of kinase activity, as well as a comparison of theeffect on blood glucose.

FIG. 5 shows impact of a compound of the invention on tumor size, andthat localization and viability of spleen-derived marginal zone B cellsis not affected by the compound, in contrast to a Pan-PI3K inhibitor.

FIG. 6 shows the in vivo efficacy of a compound of the invention againsta tumor lacking a PI3Kα mutation, compared to a kinase inhibitor withspecificity for mTor.

FIG. 7 shows the synergistic effect of combining a compound of theinvention with a MEK inhibitor.

DETAILED DESCRIPTION OF THE INVENTION

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. It is intended thatthe appended claims define the scope of the invention and that methodsand structures within the scope of these claims and their equivalents becovered thereby.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart to which this invention belongs. All patents and publicationsreferred to herein are incorporated by reference.

As used in the specification and claims, the singular form “a”, “an” and“the” includes plural references unless the context clearly dictatesotherwise.

As used herein, “agent” or “biologically active agent” refers to abiological, pharmaceutical, or chemical compound or other moiety.Non-limiting examples include simple or complex organic or inorganicmolecule, a peptide, a protein, an oligonucleotide, an antibody, anantibody derivative, antibody fragment, a vitamin derivative, acarbohydrate, a toxin, or a chemotherapeutic compound. Various compoundscan be synthesized, for example, small molecules and oligomers (e.g.,oligopeptides and oligonucleotides), and synthetic organic compoundsbased on various core structures. In addition, various natural sourcescan provide compounds for screening, such as plant or animal extracts,and the like. A skilled artisan can readily recognize that there is nolimit as to the structural nature of the agents of the presentinvention.

The term “agonist” as used herein refers to a compound having theability to initiate or enhance a biological function of a targetprotein, whether by inhibiting the activity or expression of the targetprotein. Accordingly, the term “agonist” is defined in the context ofthe biological role of the target polypeptide. While preferred agonistsherein specifically interact with (e.g., bind to) the target, compoundsthat initiate or enhance a biological activity of the target polypeptideby interacting with other members of the signal transduction pathway ofwhich the target polypeptide is a member are also specifically includedwithin this definition.

The terms “antagonist” and “inhibitor” are used interchangeably, andthey refer to a compound having the ability to inhibit a biologicalfunction of a target protein, whether by inhibiting the activity orexpression of the target protein. Accordingly, the terms “antagonist”and “inhibitors” are defined in the context of the biological role ofthe target protein. While preferred antagonists herein specificallyinteract with (e.g., bind to) the target, compounds that inhibit abiological activity of the target protein by interacting with othermembers of the signal transduction pathway of which the target proteinis a member are also specifically included within this definition. Apreferred biological activity inhibited by an antagonist is associatedwith the development, growth, or spread of a tumor, or an undesiredimmune response as manifested in autoimmune disease.

An “anti-cancer agent”, “anti-tumor agent” or “chemotherapeutic agent”refers to any agent useful in the treatment of a neoplastic condition.One class of anti-cancer agents comprises chemotherapeutic agents.“Chemotherapy” means the administration of one or more chemotherapeuticdrugs and/or other agents to a cancer patient by various methods,including intravenous, oral, intramuscular, intraperitoneal,intravesical, subcutaneous, transdermal, buccal, or inhalation or in theform of a suppository.

The term “cell proliferation” refers to a phenomenon by which the cellnumber has changed as a result of division. This term also encompassescell growth by which the cell morphology has changed (e.g., increased insize) consistent with a proliferative signal.

The terms “co-administration,” “administered in combination with,” andtheir grammatical equivalents, encompass administration of two or moreagents to an animal so that both agents and/or their metabolites arepresent in the animal at the same time. Co-administration includessimultaneous administration in separate compositions, administration atdifferent times in separate compositions, or administration in acomposition in which both agents are present.

The term “effective amount” or “therapeutically effective amount” refersto that amount of a compound described herein that is sufficient toeffect the intended application including but not limited to diseasetreatment, as defined below. The therapeutically effective amount mayvary depending upon the intended application (in vitro or in vivo), orthe subject and disease condition being treated, e.g., the weight andage of the subject, the severity of the disease condition, the manner ofadministration and the like, which can readily be determined by one ofordinary skill in the art. The term also applies to a dose that willinduce a particular response in target cells, e.g., reduction ofplatelet adhesion and/or cell migration. The specific dose will varydepending on the particular compounds chosen, the dosing regimen to befollowed, whether it is administered in combination with othercompounds, timing of administration, the tissue to which it isadministered, and the physical delivery system in which it is carried.

“Treatment”, “treating”, “palliating” and “ameliorating”, as usedherein, are used interchangeably. These terms refer to an approach forobtaining beneficial or desired results including but not limited totherapeutic benefit and/or a prophylactic benefit. By therapeuticbenefit is meant eradication or amelioration of the underlying disorderbeing treated. Also, a therapeutic benefit is achieved with theeradication or amelioration of one or more of the physiological symptomsassociated with the underlying disorder such that an improvement isobserved in the patient, notwithstanding that the patient may still beafflicted with the underlying disorder. For prophylactic benefit, thecompositions may be administered to a patient at risk of developing aparticular disease, or to a patient reporting one or more of thephysiological symptoms of a disease, even though a diagnosis of thisdisease may not have been made.

A “therapeutic effect,” as used herein, encompasses a therapeuticbenefit and/or a prophylactic benefit as described above. A prophylacticeffect includes delaying or eliminating the appearance of a disease orcondition, delaying or eliminating the onset of symptoms of a disease orcondition, slowing, halting, or reversing the progression of a diseaseor condition, or any combination thereof.

The term “pharmaceutically acceptable salt” refers to salts derived froma variety of organic and inorganic counter ions well known in the art.Pharmaceutically acceptable acid addition salts can be formed withinorganic acids and organic acids. Inorganic acids from which salts canbe derived include, for example, hydrochloric acid, hydrobromic acid,sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acidsfrom which salts can be derived include, for example, acetic acid,propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid,malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid,ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and thelike. Pharmaceutically acceptable base addition salts can be formed withinorganic and organic bases. Inorganic bases from which salts can bederived include, for example, sodium, potassium, lithium, ammonium,calcium, magnesium, iron, zinc, copper, manganese, aluminum, and thelike. Organic bases from which salts can be derived include, forexample, primary, secondary, and tertiary amines, substituted aminesincluding naturally occurring substituted amines, cyclic amines, basicion exchange resins, and the like, specifically such as isopropylamine,trimethylamine, diethylamine, triethylamine, tripropylamine, andethanolamine. In some embodiments, the pharmaceutically acceptable baseaddition salt is chosen from ammonium, potassium, sodium, calcium, andmagnesium salts.

“Pharmaceutically acceptable carrier” or “pharmaceutically acceptableexcipient” includes any and all solvents, dispersion media, coatings,antibacterial and antifungal agents, isotonic and absorption delayingagents and the like. The use of such media and agents forpharmaceutically active substances is well known in the art. Exceptinsofar as any conventional media or agent is incompatible with theactive ingredient, its use in the therapeutic compositions of theinvention is contemplated. Supplementary active ingredients can also beincorporated into the compositions.

“Signal transduction” is a process during which stimulatory orinhibitory signals are transmitted into and within a cell to elicit anintracellular response. A modulator of a signal transduction pathwayrefers to a compound which modulates the activity of one or morecellular proteins mapped to the same specific signal transductionpathway. A modulator may augment (agonist) or suppress (antagonist) theactivity of a signaling molecule.

The term “selective inhibition” or “selectively inhibit” as applied to abiologically active agent refers to the agent's ability to selectivelyreduce the target signaling activity as compared to off-target signalingactivity, via direct or interact interaction with the target.

The term “B-ALL” as used herein refers to B-cell Acute LymphoblasticLeukemia.

“Subject” refers to an animal, such as a mammal, for example a human.The methods described herein can be useful in both human therapeuticsand veterinary applications. In some embodiments, the patient is amammal, and in some embodiments, the patient is human.

“Radiation therapy” means exposing a patient, using routine methods andcompositions known to the practitioner, to radiation emitters such asalpha-particle emitting radionucleotides (e.g., actinium and thoriumradionuclides), low linear energy transfer (LET) radiation emitters(i.e., beta emitters), conversion electron emitters (e.g., strontium-89and samarium-153-EDTMP, or high-energy radiation, including withoutlimitation x-rays, gamma rays, and neutrons.

“Prodrug” is meant to indicate a compound that may be converted underphysiological conditions or by solvolysis to a biologically activecompound described herein. Thus, the term “prodrug” refers to aprecursor of a biologically active compound that is pharmaceuticallyacceptable. A prodrug may be inactive when administered to a subject,but is converted in vivo to an active compound, for example, byhydrolysis. The prodrug compound often offers advantages of solubility,tissue compatibility or delayed release in a mammalian organism (see,e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier,Amsterdam). A discussion of prodrugs is provided in Higuchi, T., et al.,“Pro-drugs as Novel Delivery Systems,” A.C.S. Symposium Series, Vol. 14,and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche,American Pharmaceutical Association and Pergamon Press, 1987, both ofwhich are incorporated in full by reference herein. The term “prodrug”is also meant to include any covalently bonded carriers, which releasethe active compound in vivo when such prodrug is administered to amammalian subject. Prodrugs of an active compound, as described herein,may be prepared by modifying functional groups present in the activecompound in such a way that the modifications are cleaved, either inroutine manipulation or in vivo, to the parent active compound. Prodrugsinclude compounds wherein a hydroxy, amino or mercapto group is bondedto any group that, when the prodrug of the active compound isadministered to a mammalian subject, cleaves to form a free hydroxy,free amino or free mercapto group, respectively. Examples of prodrugsinclude, but are not limited to, acetate, formate and benzoatederivatives of an alcohol or acetamide, formamide and benzamidederivatives of an amine functional group in the active compound and thelike.

The term “in vivo” refers to an event that takes place in a subject'sbody.

The term “in vitro” refers to an event that takes places outside of asubject's body. For example, an in vitro assay encompasses any assay runoutside of a subject assay. In vitro assays encompass cell-based assaysin which cells alive or dead are employed. In vitro assays alsoencompass a cell-free assay in which no intact cells are employed.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures wherein hydrogen is replaced by deuterium or tritium, orwherein carbon atom is replaced by ¹³C- or ¹⁴C-enriched carbon, arewithin the scope of this invention.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of atoms that constitutesuch compounds. For example, the compounds may be radiolabeled withradioactive isotopes, such as for example tritium (³H), iodine-125(¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations of the compounds ofthe present invention, whether radioactive or not, are encompassedwithin the scope of the present invention.

When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included. The term “about” when referring toa number or a numerical range means that the number or numerical rangereferred to is an approximation within experimental variability (orwithin statistical experimental error), and thus the number or numericalrange may vary from, for example, between 1% and 15% of the statednumber or numerical range. The term “comprising” (and related terms suchas “comprise” or “comprises” or “having” or “including”) includes thoseembodiments, for example, an embodiment of any composition of matter,composition, method, or process, or the like, that “consist of” or“consist essentially of” the described features.

The following abbreviations and terms have the indicated meaningsthroughout PI3-K=Phosphoinositide 3-kinase; PI=phosphatidylinositol;PDK=Phosphoinositide Dependent Kinase; DNA-PK=Deoxyribose Nucleic AcidDependent Protein Kinase; PIKK=Phosphoinositide Kinase Like Kinase;AIDS=Acquired Immuno Deficiency Syndrome; TLC=Thin Layer Chromatography;MeOH=Methanol; and CHCl₃=Chloroform.

Abbreviations used herein have their conventional meaning within thechemical and biological arts.

“Alkyl” refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, containing nounsaturation, having from one to ten carbon atoms (e.g., C₁-C₁₀ alkyl).Whenever it appears herein, a numerical range such as “1 to 10” refersto each integer in the given range; e.g., “1 to 10 carbon atoms” meansthat the alkyl group may consist of 1 carbon atom, 2 carbon atoms, 3carbon atoms, etc., up to and including 10 carbon atoms, although thepresent definition also covers the occurrence of the term “alkyl” whereno numerical range is designated. In some embodiments, it is a C₁-C₄alkyl group. Typical alkyl groups include, but are in no way limited to,methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butylisobutyl, tertiary butyl, pentyl, isopentyl, neopentyl, hexyl, septyl,octyl, nonyl, decyl, and the like. The alkyl is attached to the rest ofthe molecule by a single bond, for example, methyl (Me), ethyl (Et),n-propyl, 1-methylethyl (iso-propyl), n-butyl, n-pentyl,1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, and the like.Unless stated otherwise specifically in the specification, an alkylgroup is optionally substituted by one or more of substituents whichindependently are: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy,halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl,—OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—OC(O)N(R^(a))₂, —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a),—N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)^(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂ whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

An “alkene” moiety refers to a group consisting of at least two carbonatoms and at least one carbon-carbon double bond, and an “alkyne” moietyrefers to a group consisting of at least two carbon atoms and at leastone carbon-carbon triple bond. The alkyl moiety, whether saturated orunsaturated, may be branched, straight chain, or cyclic.

“Alkenyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one double bond, and having from two to ten carbon atoms (i.e.,C₂-C₁₀ alkenyl). Whenever it appears herein, a numerical range such as“2 to 10” refers to each integer in the given range; e.g., “2 to 10carbon atoms” means that the alkenyl group may consist of 2 carbonatoms, 3 carbon atoms, etc., up to and including 10 carbon atoms. Incertain embodiments, an alkenyl comprises two to eight carbon atoms. Inother embodiments, an alkenyl comprises two to five carbon atoms (e.g.,C₂-C₅ alkenyl). The alkenyl is attached to the rest of the molecule by asingle bond, for example, ethenyl (i.e., vinyl), prop-1-enyl (i.e.,allyl), but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like. Unlessstated otherwise specifically in the specification, an alkenyl group isoptionally substituted by one or more substituents which independentlyare: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano,trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a),—SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—OC(O)N(R^(a))₂, —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a),—N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Alkynyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one triple bond, having from two to ten carbon atoms (i.e., C₂-C₁₀alkynyl). Whenever it appears herein, a numerical range such as “2 to10” refers to each integer in the given range; e.g., “2 to 10 carbonatoms” means that the alkynyl group may consist of 2 carbon atoms, 3carbon atoms, etc., up to and including 10 carbon atoms. In certainembodiments, an alkynyl comprises two to eight carbon atoms. In otherembodiments, an alkynyl has two to five carbon atoms (e.g., C₂-C₅alkynyl). The alkynyl is attached to the rest of the molecule by asingle bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl,and the like. Unless stated otherwise specifically in the specification,an alkynyl group is optionally substituted by one or more substituentswhich independently are: alkyl, heteroalkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), SR^(a),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂,—C(O)N(R^(a))₂, —N(R^(a))C(O)O R^(a), —N(R^(a))C(O)R^(a),—N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Carboxaldehyde” refers to a —(C═O)H radical.

“Carboxyl” refers to a —(C═O)OH radical.

“Cyano” refers to a —CN radical.

“Cycloalkyl” refers to a monocyclic or polycyclic radical that containsonly carbon and hydrogen, and may be saturated, or partiallyunsaturated. Cycloalkyl groups include groups having from 3 to 10 ringatoms (i.e., C₂-C₁₀ cycloalkyl). Whenever it appears herein, a numericalrange such as “3 to 10” refers to each integer in the given range; e.g.,“3 to 10 carbon atoms” means that the cycloalkyl group may consist of 3carbon atoms, etc., up to and including 10 carbon atoms. In someembodiments, it is a C₃-C₈ cycloalkyl radical. In some embodiments, itis a C₃-C₅ cycloalkyl radical. Illustrative examples of cycloalkylgroups include, but are not limited to the following moieties:cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl,cyclohexenyl, cycloseptyl, cyclooctyl, cyclononyl, cyclodecyl,norbornyl, and the like. Unless stated otherwise specifically in thespecification, a cycloalkyl group is optionally substituted by one ormore substituents which independently are: alkyl, heteroalkyl, alkenyl,alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂,—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

The term “alkoxy” refers to the group —O-alkyl, including from 1 to 8carbon atoms of a straight, branched, cyclic configuration andcombinations thereof attached to the parent structure through an oxygen.Examples include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy,cyclohexyloxy and the like. “Lower alkoxy” refers to alkoxy groupscontaining one to six carbons. In some embodiments, C₁-C₄ alkyl, is analkyl group which encompasses both straight and branched chain alkyls offrom 1 to 4 carbon atoms.

The term “substituted alkoxy” refers to alkoxy wherein the alkylconstituent is substituted (i.e., —O-(substituted alkyl)). Unless statedotherwise specifically in the specification, the alkyl moiety of analkoxy group is optionally substituted by one or more substituents whichindependently are: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy,halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl,—OR^(a), SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—OC(O)N(R^(a))₂, —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a),—N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a)) (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

The term “alkoxycarbonyl” refers to a group of the formula(alkoxy)(C═O)— attached through the carbonyl carbon wherein the alkoxygroup has the indicated number of carbon atoms. Thus a C₁-C₆alkoxycarbonyl group is an alkoxy group having from 1 to 6 carbon atomsattached through its oxygen to a carbonyl linker. “Lower alkoxycarbonyl”refers to an alkoxycarbonyl group wherein the alkoxy group is a loweralkoxy group. In some embodiments, C₁-C₄ alkoxy, is an alkoxy groupwhich encompasses both straight and branched chain alkoxy groups of from1 to 4 carbon atoms.

The term “substituted alkoxycarbonyl” refers to the group (substitutedalkyl)-O—C(O)— wherein the group is attached to the parent structurethrough the carbonyl functionality. Unless stated otherwise specificallyin the specification, the alkyl moiety of an alkoxycarbonyl group isoptionally substituted by one or more substituents which independentlyare: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano,trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a),—SR^(a), —OC(O)—R^(a), —N(R^(a)), —C(O)R^(a), —C(O)OR^(a),—OC(O)N(R^(a))₂, —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a),—N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Acyl” refers to the groups (alkyl)-C(O)—, (aryl)-C(O)—,(heteroaryl)-C(O)—, (heteroalkyl)-C(O)—, and (heterocycloalkyl)-C(O)—,wherein the group is attached to the parent structure through thecarbonyl functionality. In some embodiments, it is a C₁-C₁₀ acyl radicalwhich refers to the total number of chain or ring atoms of the alkyl,aryl, heteroaryl or heterocycloalkyl portion of the acyloxy group plusthe carbonyl carbon of acyl, i.e three other ring or chain atoms pluscarbonyl. If the R radical is heteroaryl or heterocycloalkyl, the heteroring or chain atoms contribute to the total number of chain or ringatoms. Unless stated otherwise specifically in the specification, the“R” of an acyloxy group is optionally substituted by one or moresubstituents which independently are: alkyl, heteroalkyl, alkenyl,alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), SR^(a),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂,—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Acyloxy” refers to a R(C═O)O— radical wherein “R” is alkyl, aryl,heteroaryl, heteroalkyl, or heterocycloalkyl, which are as describedherein. In some embodiments, it is a C₁-C₄ acyloxy radical which refersto the total number of chain or ring atoms of the alkyl, aryl,heteroaryl or heterocycloalkyl portion of the acyloxy group plus thecarbonyl carbon of acyl, i.e three other ring or chain atoms pluscarbonyl. If the R radical is heteroaryl or heterocycloalkyl, the heteroring or chain atoms contribute to the total number of chain or ringatoms. Unless stated otherwise specifically in the specification, the“R” of an acyloxy group is optionally substituted by one or moresubstituents which independently are: alkyl, heteroalkyl, alkenyl,alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂,—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2-S(O)_(t)OR^(a) (where t is 1or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Amino” or “amine” refers to a —N(R^(a))₂ radical group, where eachR^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl, unless statedotherwise specifically in the specification. When a —N(R^(a))₂ group hastwo R^(a) other than hydrogen they can be combined with the nitrogenatom to form a 4-, 5-, 6-, or 7-membered ring. For example, —N(R^(a))₂is meant to include, but not be limited to, 1-pyrrolidinyl and4-morpholinyl. Unless stated otherwise specifically in thespecification, an amino group is optionally substituted by one or moresubstituents which independently are: alkyl, heteroalkyl, alkenyl,alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂,—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))C(O)N(R^(a))₂, —N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl and each of thesemoieties may be optionally substituted as defined herein.

The term “substituted amino” also refers to N-oxides of the groups—NHR^(a), and NR^(a)R^(a) each as described above. N-oxides can beprepared by treatment of the corresponding amino group with, forexample, hydrogen peroxide or m-chloroperoxybenzoic acid. The personskilled in the art is familiar with reaction conditions for carrying outthe N-oxidation.

“Amide” or “amido” refers to a chemical moiety with formula —C(O)N(R)₂or —NHC(O)R, where R is selected from the group consisting of hydrogen,alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) andheteroalicyclic (bonded through a ring carbon), each of which moiety mayitself be optionally substituted. In some embodiments it is a C₁-C₄amido or amide radical, which includes the amide carbonyl in the totalnumber of carbons in the radical. The R₂ of —N(R)₂ of the amide mayoptionally be taken together with the nitrogen to which it is attachedto form a 4-, 5-, 6-, or 7-membered ring. Unless stated otherwisespecifically in the specification, an amido group is optionallysubstituted independently by one or more of the substituents asdescribed herein for alkyl, cycloalkyl, aryl, heteroaryl, orheterocycloalkyl. An amide may be an amino acid or a peptide moleculeattached to a compound of Formula (I), thereby forming a prodrug. Anyamine, hydroxy, or carboxyl side chain on the compounds described hereincan be amidified. The procedures and specific groups to make such amidesare known to those of skill in the art and can readily be found inreference sources such as Greene and Wuts, Protective Groups in OrganicSynthesis, 3^(rd) Ed., John Wiley & Sons, New York, N.Y., 1999, which isincorporated herein by reference in its entirety.

“Aromatic” or “aryl” refers to an aromatic radical with six to ten ringatoms (e.g., C₆-C₁₀ aromatic or C₆-C₁₀ aryl) which has at least one ringhaving a conjugated pi electron system which is carbocyclic (e.g.,phenyl, fluorenyl, and naphthyl). Bivalent radicals formed fromsubstituted benzene derivatives and having the free valences at ringatoms are named as substituted phenylene radicals. Bivalent radicalsderived from univalent polycyclic hydrocarbon radicals whose names endin “-yl” by removal of one hydrogen atom from the carbon atom with thefree valence are named by adding “-idene” to the name of thecorresponding univalent radical, e.g., a naphthyl group with two pointsof attachment is termed naphthylidene. Whenever it appears herein, anumerical range such as “6 to 10” refers to each integer in the givenrange; e.g., “6 to 10 ring atoms” means that the aryl group may consistof 6 ring atoms, 7 ring atoms, etc., up to and including 10 ring atoms.The term includes monocyclic or fused-ring polycyclic (i.e., rings whichshare adjacent pairs of ring atoms) groups. Unless stated otherwisespecifically in the specification, an aryl moiety is optionallysubstituted by one or more substituents which are independently: alkyl,heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano,trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a),—SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—OC(O)N(R^(a))₂, —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a),—N(R^(a))C(O)R^(a), —N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Aralkyl” or “arylalkyl” refers to an (aryl)alkyl- radical wherein thearylalkyl moiety is attached via the alkyl portion of the moiety. Aryland alkyl are as disclosed herein and are optionally substituted by oneor more of the substituents described as suitable substituents for aryland alkyl respectively.

“Ester” refers to a chemical radical of formula —COOR, where R isselected from the group consisting of alkyl, cycloalkyl, aryl,heteroaryl (bonded through a ring carbon) and heteroalicyclic (bondedthrough a ring carbon). Any amine, hydroxy, or carboxyl side chain onthe compounds described herein can be esterified. The procedures andspecific groups to make such esters are known to those of skill in theart and can readily be found in reference sources such as Greene andWuts, Protective Groups in Organic Synthesis, 3^(rd) Ed., John Wiley &Sons, New York, N.Y., 1999, which is incorporated herein by reference inits entirety. Unless stated otherwise specifically in the specification,an ester group is optionally substituted by one or more substituentswhich independently are: alkyl, heteroalkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR^(a), —SR^(a),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —OC(O)N(R^(a))₂,—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), N(R^(a))C(O)R^(a),—N(R^(a))C(O)N(R^(a))₂, N(R^(a))C(NR^(a))N(R^(a))₂,—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or 2), or PO₃(R^(a))₂, whereeach R^(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

“Fluoroalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more fluoro radicals, as defined above, forexample, trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl,1-fluoromethyl-2-fluoroethyl, and the like. The alkyl part of thefluoroalkyl radical may be optionally substituted as defined above foran alkyl group.

“Halo”, “halide”, or, alternatively, “halogen” means fluoro, chloro,bromo or iodo. The terms “haloalkyl,” “haloalkenyl,” “haloalkynyl” and“haloalkoxy” include alkyl, alkenyl, alkynyl and alkoxy structures thatare substituted with one or more halo groups or with combinationsthereof. For example, the terms “fluoroalkyl” and “fluoroalkoxy” includehaloalkyl and haloalkoxy groups, respectively, in which the halo isfluorine.

“Heteroalkyl”, “heteroalkenyl” and “heteroalkynyl” include optionallysubstituted alkyl, alkenyl and alkynyl radicals and which have one ormore skeletal chain atoms selected from an atom other than carbon, e.g.,oxygen, nitrogen, sulfur, phosphorus or combinations thereof. Anumerical range may be given, e.g., C₁-C₄ heteroalkyl which refers tothe chain length in total which in this example is 4 atoms long. Forexample, a —CH₂OCH₂CH₃ radical is referred to as a “C₄” heteroalkyl,which includes the heteroatom center in the atom chain lengthdescription. Connection to the rest of the molecule may be througheither a heteroatom or a carbon in the heteroalkyl chain. A heteroalkylgroup may be substituted with one or more substituents whichindependently are: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy,halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a), —SR^(a),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where tis 1 or 2), —S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂(where t is 1 or 2), or PO₃(R^(a))₂, where each R^(a) is independentlyhydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl,aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl orheteroarylalkyl.

“Heteroalkylaryl” refers “to an -(heteroalkyl)aryl radical whereheteroalkyl and aryl are as disclosed herein and which are optionallysubstituted by one or more of the substituents described as suitablesubstituents for heteroalkyl and aryl respectively.

“Heteroaryl” or, alternatively, “heteroaromatic” refers to a 5- to18-membered aromatic radical (e.g., C₅-C₁₃ heteroaryl) that includes oneor more ring heteroatoms selected from nitrogen, oxygen and sulfur, andwhich may be a monocyclic, bicyclic, tricyclic or tetracyclic ringsystem. Whenever it appears herein, a numerical range such as “5 to 18”refers to each integer in the given range; e.g., “5 to 18 ring atoms”means that the heteroaryl group may consist of 5 ring atoms, 6 ringatoms, etc., up to and including 18 ring atoms. Bivalent radicalsderived from univalent heteroaryl radicals whose names end in “-yl” byremoval of one hydrogen atom from the atom with the free valence arenamed by adding “-idene” to the name of the corresponding univalentradical, e.g., a pyridyl group with two points of attachment is apyridylidene. An N-containing “heteroaromatic” or “heteroaryl” moietyrefers to an aromatic group in which at least one of the skeletal atomsof the ring is a nitrogen atom. The polycyclic heteroaryl group may befused or non-fused. The heteroatom(s) in the heteroaryl radical isoptionally oxidized. One or more nitrogen atoms, if present, areoptionally quaternized. The heteroaryl is attached to the rest of themolecule through any atom of the ring(s). Examples of heteroarylsinclude, but are not limited to, azepinyl, acridinyl, benzimidazolyl,benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl,benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl,benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl,benzoxazolyl, benzodioxolyl, benzodioxinyl, benzoxazolyl, benzopyranyl,benzopyranonyl, benzofuranyl, benzofuranonyl, benzofurazanyl,benzothiazolyl, benzothienyl (benzothiophenyl),benzothieno[3,2-d]pyrimidinyl, benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,cyclopenta[d]pyrimidinyl,6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl,6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl,dihenzothiophenyl, furanyl, furazanyl, furanonyl, furo[3,2-c]pyridinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl,indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl,isoquinolyl, indolizinyl, isoxazolyl,5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl,1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl,5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl,phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl,purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl,pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl,pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl,quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl,5,6,7,8-tetrahydroquinazolinyl,5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl.6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl,thiapyranyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl,thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e.thienyl). Unless stated otherwise specifically in the specification, aheteroaryl moiety is optionally substituted by one or more substituentswhich are independently: alkyl, heteroalkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo,trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a),—N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂ (where t is 1 or2), or PO₃(R^(a))₂, where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl,heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

Substituted heteroaryl also includes ring systems substituted with oneor more oxide (—O—) substituents, such as pyridinyl N-oxides.

“Heteroarylalkyl” refers to a moiety having a heteroaryl moiety, asdescribed herein, connected to an alkyl moiety, as described herein,wherein the connection to the remainder of the molecule is through thealkyl group. Heteroaryl and alkyl are as disclosed herein and areoptionally substituted by one or more of the substituents described assuitable substituents for heteroaryl and alkyl respectively.

“Heterocycloalkyl” refers to a stable 3- to 18-membered non-aromaticring radical that comprises two to twelve carbon atoms and from one tosix heteroatoms selected from nitrogen, oxygen and sulfur. Whenever itappears herein, a numerical range such as “3 to 18” refers to eachinteger in the given range; e.g., “3 to 18 ring atoms” means that theheterocycloalkyl group may consist of 3 ring atoms, 4 ring atoms, etc.,up to and including 18 ring atoms. In some embodiments, it is a C₅-C₁₀heterocycloalkyl. In some embodiments, it is a C₄-C₁₀ heterocycloalkyl.In some embodiments, it is a C₃-C₁₀ heterocycloalkyl. Unless statedotherwise specifically in the specification, the heterocycloalkylradical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system,which may include fused or bridged ring systems. The heteroatoms in theheterocycloalkyl radical may be optionally oxidized. One or morenitrogen atoms, if present, are optionally quaternized. Theheterocycloalkyl radical is partially or fully saturated. Theheterocycloalkyl may be attached to the rest of the molecule through anyatom of the ring(s). Examples of such heterocycloalkyl radicals include,but are not limited to, dioxolanyl, thienyl[1,3]dithianyl,decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl,isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl,2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl,piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl,quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl,tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl,1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless statedotherwise specifically in the specification, a heterocycloalkyl moietyis optionally substituted by one or more substituents whichindependently are: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy,halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a), —SR^(a),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where tis 1 or 2), —S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)N(R^(a))₂(where t is 1 or 2), or PO₃(R^(a))₂, where each R^(a) is independentlyhydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl,aralkyl, heterocycloalkyl, heteroaryl or heteroarylalkyl.

“Heterocycloalkyl” also includes bicyclic ring systems wherein onenon-aromatic ring, usually with 3 to 7 ring atoms, contains at least 2carbon atoms in addition to 1-3 heteroatoms independently selected fromoxygen, sulfur, and nitrogen, as well as combinations comprising atleast one of the foregoing heteroatoms; and the other ring, usually with3 to 7 ring atoms, optionally contains 1-3 heteroatoms independentlyselected from oxygen, sulfur, and nitrogen and is not aromatic.

“Heterocycloalkyloxy” refers to a (heterocycloalkyl)-O— moiety, wherethe heterocycloalkyl moiety is attached via a carbon atom to oxygen,wherein the oxygen functions as a linker to attach the moiety to acompound. The heterocycloalkyl is as described herein and is optionallysubstituted by one or more substituents described herein as suitable forheterocycloalkyl.

“Isomers” are different compounds that have the same molecular formula.“Stereoisomers” are isomers that differ only in the way the atoms arearranged in space. “Enantiomers” are a pair of stereoisomers that arenon-superimposable mirror images of each other. A 1:1 mixture of a pairof enantiomers is a “racemic” mixture. The term “(±)” is used todesignate a racemic mixture where appropriate. “Diastereoisomers” arestereoisomers that have at least two asymmetric atoms, but which are notmirror-images of each other. The absolute stereochemistry is specifiedaccording to the Cahn-Ingold-Prelog R-S system. When a compound is apure enantiomer the stereochemistry at each chiral carbon can bespecified by either R or S. Resolved compounds whose absoluteconfiguration is unknown can be designated (+) or (−) depending on thedirection (dextro- or levorotatory) which they rotate plane polarizedlight at the wavelength of the sodium D line. Certain of the compoundsdescribed herein contain one or more asymmetric centers and can thusgive rise to enantiomers, diastereomers, and other stereoisomeric formsthat can be defined, in terms of absolute stereochemistry, as (R)- or(S)-. The present chemical entities, pharmaceutical compositions andmethods are meant to include all such possible isomers, includingracemic mixtures, optically pure forms and intermediate mixtures.Optically active (R)- and (S)-isomers can be prepared using chiralsynthons or chiral reagents, or resolved using conventional techniques.The optical activity of a compound can be analyzed via any suitablemethod, including but not limited to chiral chromatography andpolarimetry, and the degree of predominance of one stereoisomer over theother isomer can be determined.

When the compounds described herein contain olefinic double bonds orother centers of geometric asymmetry, and unless specified otherwise, itis intended that the compounds include both E and Z geometric isomers.

“Moiety” refers to a specific segment or functional group of a molecule.Chemical moieties are often recognized chemical entities embedded in orappended to a molecule.

“Nitro” refers to the —NO₂ radical.

“Oxa” refers to the —O— radical.

“Oxo” refers to the ═O radical.

“Tautomers” are structurally distinct isomers that interconvert bytautomerization. “Tautomerization” is a form of isomerization andincludes prototropic or proton-shift tautomerization, which isconsidered a subset of acid-base chemistry. “Prototropictautomerization” or “proton-shift tautomerization” involves themigration of a proton accompanied by changes in bond order, often theinterchange of a single bond with an adjacent double bond. Wheretautomerization is possible (e.g., in solution), a chemical equilibriumof tautomers can be reached. An example of tautomerization is keto-enoltautomerization. A specific example of keto-enol tautomerization is theinterconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-onetautomers. Another example of tautomerization is phenol-ketotautomerization. A specific example of phenol-keto tautomerization isthe interconversion of pyridin-4-ol and pyridin-4(1H)-one tautomers.

The compounds or the present invention may also contain unnaturalproportions or atomic isotopes at one or more of atoms that constitutesuch compounds. For example, the compounds may be radiolabeled withradioactive isotopes, such as for example tritium (³H), iodine-125(¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations of the compounds ofthe present invention, whether radioactive or not, are encompassedwithin the scope of the present invention.

A “leaving group or atom” is any group or atom that will, under thereaction conditions, cleave from the starting material, thus promotingreaction at a specified site. Suitable examples of such groups unlessotherwise specified are halogen atoms, mesyloxy,p-nitrobenzensulphonyloxy and tosyloxy groups.

“Protecting group” has the meaning conventionally associated with it inorganic synthesis, i.e. a group that selectively blocks one or morereactive sites in a multifunctional compound such that a chemicalreaction can be carried out selectively on another unprotected reactivesite and such that the group can readily be removed after the selectivereaction is complete. A variety of protecting groups are disclosed, forexample, in T. H. Greene and P. G. M. Wuts, Protective Groups in OrganicSynthesis, Third Edition. John Wiley & Sons, New York (1999). Forexample, a hydroxy protected form is where at least one of the hydroxygroups present in a compound is protected with a hydroxy protectinggroup. Likewise, amines and other reactive groups may similarly beprotected.

“Solvate” refers to a compound (e.g., a compound selected from Formula Ior a pharmaceutically acceptable salt thereof) in physical associationwith one or more molecules of a pharmaceutically acceptable solvent. Itwill be understood that “a compound or Formula I” encompass the compoundor Formula I and solvates of the compound, as well as mixtures thereof.

“Substituted” means that the referenced group may be substituted withone or more additional group(s) individually and independently selectedfrom acyl, alkyl, alkylaryl, cycloalkyl, aralkyl, aryl, carbohydrate,carbonate, heteroaryl, heterocycloalkyl, hydroxy, alkoxy, aryloxy,mercapto, alkylthio, arylthio, cyano, halo, carbonyl, ester,thiocarbonyl, isocyanato, thiocyanato, isothiocyanato, nitro, oxo,perhaloalkyl, perfluoroalkyl, phosphate, silyl, sulfinyl, sulfonyl,sulfonamidyl, sulfoxyl, sulfonate, urea, and amino, including mono- anddi-substituted amino groups, and the protected derivatives thereof.Di-substituted amino groups encompass those which form a ring togetherwith the nitrogen of the amino group, such as for instance, morpholino.The substituents themselves may be substituted, for example, a cycloakylsubstituent may have a halide substituted at one or more ring carbons,and the like. The protecting groups that may form the protectivederivatives of the above substituents are known to those of skill in theart and may be found in references such as Greene and Wuts, above.

“Sulfonyl” refers to the groups: —S(O₂)—H, —S(O₂)-(optionallysubstituted alkyl), —S(O₂)-(optionally substituted amino),—S(O₂)-(optionally substituted aryl), —S(O₂)-(optionally substitutedheteroaryl), and —S(O₂)-(optionally substituted heterocycloalkyl).

“Sulfonamidyl” or “sulfonamido” refers to a —S(═O)₂—NRR radical, whereeach R is selected independently from the group consisting of hydrogen,alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) andheteroalicyclic (bonded through a ring carbon). The R groups in —NRR ofthe —S(═O)₂—NRR radical may be taken together with the nitrogen to whichit is attached to form a 4-, 5-, 6-, or 7-membered ring. In someembodiments, it is a C₁-C₁₀ sulfonamido, wherein each R in sulfonamidocontains 1 carbon, 2 carbons, 3 carbons, or 4 carbons total. Asulfonamido group is optionally substituted by one or more of thesubstituents described for alkyl, cycloalkyl, aryl, heteroarylrespectively

“Sulfoxyl” refers to a —S(═O)₂OH radical.

“Sulfonate” refers to a —S(═O)₂—OR radical, where R is selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded througha ring carbon) and heteroalicyclic (bonded through a ring carbon). Asulfonate group is optionally substituted on R by one or more of thesubstituents described for alkyl, cycloalkyl aryl, heteroarylrespectively.

Where substituent groups are specified by their conventional chemicalformulae, written from left to right, they equally encompass thechemically identical substituents that would result from writing thestructure from right to left, e.g., —CH₂O— is equivalent to —OCH₂—.

Compounds of the present invention also include crystalline andamorphous forms of those compounds, including, for example, polymorphs,pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (includinganhydrates), conformational polymorphs, and amorphous forms of thecompounds, as well as mixtures thereof. “Crystalline form,” “polymorph,”and “novel form” may be used interchangeably herein, and are meant toinclude all crystalline and amorphous forms of the compound, including,for example, polymorphs, pseudopolymorphs, solvates, hydrates,unsolvated polymorphs (including anhydrates), conformational polymorphs,and amorphous forms, as well as mixtures thereof, unless a particularcrystalline or amorphous form is referred to.

“Solvent,” “organic solvent,” and “inert solvent” each means a solventinert under the conditions of the reaction being described inconjunction therewith including, for example, benzene, toluene,acetonitrile, tetrahydrofuran (“THF”), dimethylformamide (“DMF”),chloroform, methylene chloride (or dichloromethane), diethyl ether,methanol, N-methylpyrrolidone (“NMP”), pyridine and the like. Unlessspecified to the contrary, the solvents used in the reactions describedherein are inert organic solvents. Unless specified to the contrary, foreach gram of the limiting reagent, one cc (or mL) of solvent constitutesa volume equivalent.

Isolation and purification of the chemical entities and intermediatesdescribed herein can be effected, if desired, by any suitable separationor purification procedure such as, for example, filtration, extraction,crystallization, column chromatography, thin-layer chromatography orthick-layer chromatography, or a combination of these procedures.Specific illustrations of suitable separation and isolation procedurescan be had by reference to the examples hereinbelow. However, otherequivalent separation or isolation procedures can also be used.

When desired, the (R)- and (S)-isomers of the compounds of the presentinvention, if present, may be resolved by methods known to those skilledin the art, for example by formation of diastereoisomeric salts orcomplexes which may be separated, for example, by crystallization; viaformation of diastereoisomeric derivatives which may be separated, forexample, by crystallization, gas-liquid or liquid chromatography;selective reaction of one enantiomer with an enantiomer-specificreagent, for example enzymatic oxidation or reduction, followed byseparation of the modified and unmodified enantiomers; or gas-liquid orliquid chromatography in a chiral environment, for example on a chiralsupport, such as silica with a bound chiral ligand or in the presence ofa chiral solvent. Alternatively, a specific enantiomer may besynthesized by asymmetric synthesis using optically active reagents,substrates, catalysts or solvents, or by converting one enantiomer tothe other by asymmetric transformation.

The compounds described herein can be optionally contacted with apharmaceutically acceptable acid to form the corresponding acid additionsalts. Pharmaceutically acceptable forms of the compounds recited hereininclude pharmaceutically acceptable salts, chelates, non-covalentcomplexes, prodrugs, and mixtures thereof. In certain embodiments, thecompounds described herein are in the form of pharmaceuticallyacceptable salts. In addition, if the compound described herein isobtained as an acid addition salt, the free base can be obtained bybasifying a solution of the acid salt. Conversely, if the product is afree base, an addition salt, particularly a pharmaceutically acceptableaddition salt, may be produced by dissolving the free base in a suitableorganic solvent and treating the solution with an acid, in accordancewith conventional procedures for preparing acid addition salts from basecompounds. Those skilled in the art will recognize various syntheticmethodologies that may be used to prepare non-toxic pharmaceuticallyacceptable addition salts.

As noted above, the present invention provides various compounds thatare useful as antagonists for one or more lipid kinases and/or proteinkinases.

In one aspect, the present invention provides a compound of Formula I:

or its pharmaceutically acceptable salts thereof, wherein:

X is O or S or N;

-   -   W¹ is N, NR³, CR³, or C═O, W² is N, NR⁴, CR⁴, or C═O, W³ is N,        NR⁵ or CR⁵, W⁴ is N, C═O or CR⁶, wherein no more than two N        atoms and no more than two C═O groups are adjacent;    -   W⁵ is N or CR⁷;    -   W⁶ is N or CR⁸;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   or R³ and R⁴ taken together form a cyclic moiety; and    -   R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, alkyl,        heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,        aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,        heterocycloalkyloxy, amido, amino, acyl, acyloxy,        alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,        phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are taken        together with nitrogen to form a cyclic moiety.

In some embodiments, the compound of Formula I exists as a tautomer, andsuch tautomers are contemplated by the present invention.

In some embodiments, the compound of Formula I has the formula:

For example, a compound of Formula I is:

In some embodiments of the compound of Formula I, W¹ is CR³, W² is CR⁴,W³ is CR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸; W¹ is N, W² is CR⁴, W³ isCR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸; or W¹ is CR³, W² is N, W³ isCR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸. Formulas for such embodimentsare shown below.

In some embodiments, X is O. In other embodiments, X is S.

In some embodiments, R¹ is hydrogen. In other embodiments, R¹ is alkyl,heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy,amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,hydroxy, nitro, phosphate, urea, carbonate, or NR′R″, wherein R′ and R″are taken together with nitrogen to form a cyclic moiety.

In some embodiments, R² is hydrogen. In other embodiments, R² is, forexample, unsubstituted or substituted alkyl (including but not limitedto CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, and heptyl). In other embodiments, R² is unsubstituted orsubstituted alkenyl (including but not limited to unsubstituted orsubstituted C₂-C₅alkenyl such as, for example, vinyl, allyl, 1-methylpropen-1-yl, butenyl, or pentenyl) or unsubstituted or substitutedalkynyl (including but not limited to unsubstituted or substitutedC₂-C₅alkynyl such as acetylenyl, propargyl, butynyl, or pentynyl).Alternatively, R² is unsubstituted or substituted aryl (including butnot limited to monocyclic or bicyclic aryl) or unsubstituted orsubstituted arylalkyl (including but not limited to monocyclic orbicyclic aryl linked to alkyl wherein alkyl includes but is not limitedto CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl).In some other embodiments, R² is unsubstituted or substitutedheteroaryl, including but not limited to monocyclic and bicyclicheteroaryl. Monocyclic heteroaryl R² includes but is not limited topyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl, pyrazinyl,pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl. Bicyclicheteroaryl R² includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl. The presentinvention also provides compounds wherein R² is unsubstituted orsubstituted heteroarylalkyl, including but not limited to monocyclic andbicyclic heteroaryl as described above, that are linked to alkyl, whichin turn includes but is not limited to CH₃, —CH₂CH₃, n-propyl,isopropyl, n-butyl, sec-butyl, and pentyl. In some embodiments, R² isunsubstituted or substituted cycloalkyl (including but not limited tocyclopropyl, cyclobutyl, and cyclopentyl) or unsubstituted orsubstituted heteroalkyl (non-limiting examples include ethoxymethyl,methoxymethyl, and diethylaminomethyl). In some further embodiments, R²is unsubstituted or substituted heterocycloalkyl which includes but isnot limited to pyrrolidinyl, tetrahydrofuranyl, piperidinyl,tetrahydropyranyl, thiazolidinyl, imidazolidinyl, morpholinyl, andpiperazinyl. In yet other embodiments of the compounds of Formula I, R²is unsubstituted or substituted alkoxy including but not limited toC₁-C₄alkoxy such as methoxy, ethoxy, propoxy or butoxy. R² can also beunsubstituted or substituted heterocycloalkyloxy, including but notlimited to 4-NH piperidin-1-yl-oxy, 4-methyl piperidin-1-yl-oxy, 4-ethylpiperidin-1-yl-oxy, 4-isopropyl-piperidin-1-yl-oxy, andpyrrolidin-3-yl-oxy. In other embodiments, R² is unsubstituted orsubstituted amino, wherein the substituted amino includes but is notlimited to dimethylamino, diethylamino, di-isopropyl amino, N-methylN-ethyl amino, and dibutylamino. In some embodiments, R² isunsubstituted or substituted acyl, unsubstituted or substituted acyloxy,unsubstituted or substituted C₁-C₄acyloxy, unsubstituted or substitutedalkoxycarbonyl, unsubstituted or substituted amido, or unsubstituted orsubstituted sulfonamido. In other embodiments, R² is halo, which is —I,—F, —Cl, or —Br. In some embodiments, R² is selected from the groupconsisting of cyano, hydroxy, nitro, phosphate, urea, and carbonate.Also contemplated are R² being —CH₃, —CH₂CH₃, n-propyl, isopropyl,n-butyl, tert-butyl, sec-butyl, pentyl, hexyl, heptyl, —OCH₃, —OCH₂CH₃,or —CF₃.

In some embodiments of the compound of Formula I, W¹ is CR³. R³ can be,for example, hydrogen, unsubstituted or substituted alkyl (including butnot limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl,sec-butyl, pentyl, hexyl, and heptyl). In other embodiments, R³ isunsubstituted or substituted alkenyl (including but not limited tounsubstituted or substituted C₂-C₅alkenyl such as, for example, vinyl,allyl, 1-methyl propen-1-yl, butenyl, or pentenyl) or unsubstituted orsubstituted alkynyl (including but not limited to unsubstituted orsubstituted C₂-C₅alkynyl such as acetylenyl, propargyl, butynyl, orpentynyl). Alternatively, R³ is unsubstituted or substituted aryl(including but not limited to monocyclic or bicyclic aryl) orunsubstituted or substituted arylalkyl (including but not limited tomonocyclic or bicyclic aryl linked to alkyl wherein alkyl includes butis not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl,and pentyl). In some other embodiments, R³ is unsubstituted orsubstituted heteroaryl, including but not limited to monocyclic andbicyclic heteroaryl. Monocyclic heteroaryl R³ includes but is notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl.Bicyclic heteroaryl R³ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl. The presentinvention also provides compounds of Formula I wherein R³ isunsubstituted or substituted heteroarylalkyl, including but not limitedto monocyclic and bicyclic heteroaryl as described above, that arelinked to alkyl, which in turn includes but is not limited to CH₃,—CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl. In someembodiments, R³ is unsubstituted or substituted cycloalkyl (includingbut not limited to cyclopropyl, cyclobutyl, and cyclopentyl) orunsubstituted or substituted heteroalkyl (non-limiting examples includeethoxymethyl, methoxymethyl, and diethylaminomethyl). In some furtherembodiments, R³ is unsubstituted or substituted heterocycloalkyl whichincludes but is not limited to pyrrolidinyl, tetrahydrofuranyl,piperidinyl, tetrahydropyranyl, thiazolidinyl, imidazolidinyl,morpholinyl, and piperazinyl. In yet other embodiments of the compoundsof Formula I, R³ is unsubstituted or substituted alkoxy including butnot limited to C₁-C₄alkoxy such as methoxy, ethoxy, propoxy or butoxy.R³ can also be unsubstituted or substituted heterocycloalkyloxy,including but not limited to 4-NH piperidin-1-yl-oxy, 4-methylpiperidin-1-yl-oxy, 4-ethyl piperidin-1-yl-oxy,4-isopropyl-piperidin-1-yl-oxy, and pyrrolidin-3-yl-oxy. In otherembodiments, R³ is unsubstituted or substituted amino, wherein thesubstituted amino includes but is not limited to dimethylamino,diethylamino, di-isopropyl amino, N-methyl N-ethyl amino, anddibutylamino. In some embodiments, R³ is unsubstituted or substitutedacyl, unsubstituted or substituted acyloxy, unsubstituted or substitutedC₁-C₄acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstitutedor substituted amido, or unsubstituted or substituted sulfonamido. Inother embodiments, R³ is halo, which is —I, —F, —Cl, or —Br. In someembodiments, R³ is selected from the group consisting of cyano, hydroxy,nitro, phosphate, urea, and carbonate. Also contemplated are R³ being—CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, heptyl, —OCH₃, —OCH₂CH₃, or —CF₃.

R³ of the compounds of Formula I, can also be NR′R″ wherein R′ and R″are taken together with the nitrogen to form a cyclic moiety having from3 to 8 ring atoms. The cyclic moiety so formed may further include oneor more heteroatoms which are selected from the group consisting of S,O, and N. The cyclic moiety so formed is unsubstituted or substituted,including but not limited to morpholinyl, azetidinyl, pyrrolidinyl,piperidinyl, piperazinyl, isothiazolidinyl 1,2, dioxide, andthiomorpholinyl. Further non-limiting exemplary cyclic moieites are thefollowing:

The invention also provides compounds of Formula I, wherein when R³ is amember of the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl,heteroalkyl, heterocycloalkyl, heterocycloalkyloxy, aryl, arylalkyl,heteroaryl, heteroarylalkyl, acyl, alkoxy, amido, amino, sulfonamido,acyloxy, alkoxycarbonyl, and NR′R″ (wherein R′ and R″ are taken togetherwith nitrogen to form a cyclic moiety), then R³ is optionallysubstituted with one or more of the following substituents: alkyl,alkenyl, alkynyl, cycloalkyl, heteroalkyl, heterocycloalkyl,heterocycloalkyloxy, aryl, arylalkyl, heteroaryl, heteroarylalkyl, acyl,heterocycloalkyloxy, alkoxy, amido, amino, sulfonamido, acyloxy,alkoxycarbonyl, halo, cyano, hydroxy, nitro, phosphate, urea, carbonate,or NR′R″ wherein R′ and R″ are taken together with nitrogen to form acyclic moiety. Each of the above substituents may be further substitutedwith one or more substituents chosen from the group consisting of alkyl,alkoxy, amido, amino, sulfonamido, acyloxy, alkoxycarbonyl, halo, cyano,hydroxy, nitro, oxo, phosphate, urea, and carbonate.

For example, the invention provides compounds wherein when R³ is alkyl,the alkyl is substituted with NR′R″ wherein R′ and R″ are taken togetherwith the nitrogen to form a cyclic moiety. The cyclic moiety so formedcan be unsubstituted or substituted. Non-limiting exemplary cyclicmoieties includes but are not limited to morpholinyl, azetidinyl,pyrrolidinyl, piperidinyl, piperazinyl, and thiomorpholinyl. In otherexamples of the compounds of Formula I, when R³ is alkyl, the alkyl issubstituted with heterocycloalkyl, which includes oxetanyl, azetidinyl,tetrahydrofuranyl, pyrrolyl, tetrahydropyranyl, piperidinyl,morpholinyl, and piperazinyl. All of the above listed heterocycloaklylsubstituents can be unsubstituted or substituted.

In yet other examples of the compounds of Formula I, when R³ is alkyl,the alkyl is substituted with a 5, 6, 7, 8, 9, or 10 membered monocyclicor bicyclic heteroaryl, which is unsubstituted or substituted. Themonocyclic heteroaryl includes but is not limited to pyrrolyl, thienyl,furyl, pyridinyl, pyranyl, pyrimidinyl, pyrazinyl, pyridazinyl,imidazolyl, thiazolyl, pyrazolyl, and oxazolyl. The bicyclic heteroarylincludes but is not limited to benzothiophenyl, benzofuryl, indolyl,quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, benzothiazolyl,quinazolinyl, azaindolyl, pyrazolopyrimidinyl, purinyl, pyrrolo [1,2-b]pyridazinyl, pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl,imidazo[1, 2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

In other embodiments of the compound of Formula I, R³ is —NHR^(3′),—N(CH₃)R^(3′), —N(CH₂CH₃)R^(3′), —N(CH(CH₃)₂)R^(3′), or —OR^(3′),wherein R^(3′) is unsubstituted or substituted heterocycloalkyl(nonlimiting examples thereof include 4-NH piperidin-1-yl, 4-methylpiperidin-1-yl, 4-ethyl piperidin-1-yl, 4-isopropyl-piperidin-1-yl, andpyrrolidin-3-yl), unsubstituted or substituted monocyclic aryl, orunsubstituted or substituted monocyclic heteroaryl (including but notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl).In one example, R³ is O-aryl, i.e. phenoxy. In another example, R³ is—O-(4-methyl)piperidin-1-yl or —O-(4-isopropyl)piperidin-1-yl.

In some embodiments of the compound of Formula I, R³ is one of thefollowing moieties:

In some embodiments of the compound of Formula I, W¹ is NR³, wherein R³is hydrogen, unsubstituted or substituted C₁-C₁₀alkyl (which includesbut is not limited to —CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyltert-butyl, sec-butyl, pentyl, hexyl, and heptyl), or unsubstituted orsubstituted C₃-C₇cycloalkyl (which includes but is not limited tocyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl). In otherembodiments of the compound of Formula I, R³ is unsubstituted orsubstituted heterocycloalkyl (which includes but is not limited tooxetanyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl,piperidinyl, and piperazinyl), or unsubstituted or substitutedC₂-C₁₀heteroalkyl (which includes but is not limited to methoxyethoxy,methoxymethyl, and diethylaminoethyl). Alternatively, R³ isunsubstituted or substituted monocyclic heteroaryl (which includes butis not limited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl,pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl,and oxazolyl) or unsubstituted or substituted monocyclic aryl.

In still other embodiments, W¹ is C═O.

In some embodiments of the compound of Formula I. W² is CR⁴. R⁴ can be,for example, hydrogen, or unsubstituted or substituted alkyl (includingbut not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl,tert-butyl, sec-butyl, pentyl, hexyl, and heptyl). In other embodiments,R⁴ is unsubstituted or substituted alkenyl (including but not limited tounsubstituted or substituted C₂-C₅alkenyl such as, for example, vinyl,allyl, 1-methyl propen-1-yl, butenyl, or pentenyl) or unsubstituted orsubstituted alkynyl (including but not limited to unsubstituted orsubstituted C₂-C₅alkynyl such as acetylenyl, propargyl, butynyl, orpentynyl). Alternatively, R⁴ is unsubstituted or substituted aryl(including but not limited to monocyclic or bicyclic aryl) orunsubstituted or substituted arylalkyl (including but not limited tomonocyclic or bicyclic aryl linked to alkyl wherein alkyl includes butis not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl,and pentyl). In some other embodiments, R⁴ is unsubstituted orsubstituted heteroaryl, including but not limited to monocyclic andbicyclic heteroaryl. Monocyclic heteroaryl R⁴ includes but is notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl.Bicyclic heteroaryl R⁴ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

The present invention also provides compounds of Formula I wherein R⁴ isunsubstituted or substituted heteroarylalkyl, including but not limitedto monocyclic and bicyclic heteroaryl as described above, that arelinked to alkyl, which in turn includes but is not limited to CH₃,—CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl. In someembodiments, R⁴ is unsubstituted or substituted cycloalkyl (includingbut not limited to cyclopropyl, cyclobutyl, and cyclopentyl) orunsubstituted or substituted heteroalkyl (non-limiting examples includeethoxymethyl, methoxymethyl, and diethylaminomethyl). In some furtherembodiments, R⁴ is unsubstituted or substituted heterocycloalkyl whichincludes but is not limited to pyrrolidinyl, tetrahydrofuranyl,piperidinyl, tetrahydropyranyl, thiazolidinyl, imidazolidinyl,morpholinyl, and piperazinyl. In yet other embodiments of the compoundsof Formula I, R⁴ is unsubstituted or substituted alkoxy including butnot limited to C₁-C₄alkoxy such as methoxy, ethoxy, propoxy or butoxy.R⁴ can also be unsubstituted or substituted heterocycloalkyloxy,including but not limited to 4-NH piperidin-1-yl-oxy, 4-methylpiperidin-1-yl-oxy, 4-ethyl piperidin-1-yl-oxy,4-isopropyl-piperidin-1-yl-oxy, and pyrrolidin-3-yl-oxy. In otherembodiments, R⁴ is unsubstituted or substituted amino, wherein thesubstituted amino includes but is not limited to dimethylamino,diethylamino, di-isopropyl amino, N-methyl N-ethyl amino, anddibutylamino. In some embodiments, R⁴ is unsubstituted or substitutedacyl, unsubstituted or substituted acyloxy, unsubstituted or substitutedC₁-C₄acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstitutedor substituted amido, or unsubstituted or substituted sulfonamido. Insome embodiments, R⁴ is halo, which is —I, —F, —Cl, or —Br. In someembodiments, R⁴ is selected from the group consisting of cyano, hydroxy,nitro, phosphate, urea, or carbonate. Also contemplated are R⁴ being—CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, heptyl, —OCH₃, —OCH₂CH₃, or —CF₃.

R⁴ of the compounds of Formula I, can also be NR′R″ wherein R′ and R″are taken together with the nitrogen to form a cyclic moiety having from3 to 8 ring atoms. The cyclic moiety so formed may further include oneor more heteroatoms which are selected from the group consisting of S,O, and N. The cyclic moiety so formed is unsubstituted or substituted,including but not limited to morpholinyl, azetidinyl, pyrrolidinyl,piperidinyl, piperazinyl, isothiazolidinyl 1,2, dioxide, andthiomorpholinyl. Further non-limiting exemplary cyclic moieties are thefollowing:

The invention also provides compounds of Formula I, wherein when R⁴ is amember of the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl,heteroalkyl, heterocycloalkyl, heterocycloalkyloxy, aryl, arylalkyl,heteroaryl, heteroarylalkyl, acyl, alkoxy, amido, amino, sulfonamido,acyloxy, alkoxycarbonyl, and NR′R″ (wherein R′ and R″ are taken togetherwith nitrogen to form a cyclic moiety), then R⁴ is optionallysubstituted with one or more of the following substituents: alkyl,alkenyl, alkynyl, cycloalkyl, heteroalkyl, heterocycloalkyl,heterocycloalkyloxy, aryl, arylalkyl, heteroaryl, heteroarylalkyl, acyl,alkoxy, amido, amino, sulfonamido, acyloxy, alkoxycarbonyl, halo, cyano,hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′ and R″are taken together with nitrogen to form a cyclic moiety. Each of theabove substituents may be further substituted with one or moresubstituents chosen from the group consisting of alkyl, alkoxy, amido,amino, sulfonamido, acyloxy, alkoxycarbonyl, halo, cyano, hydroxy,nitro, oxo, phosphate, urea, and carbonate.

For example, the invention provides compounds wherein when R⁴ is alkyl,the alkyl is substituted with NR′R″ wherein R′ and R″ are taken togetherwith the nitrogen to form a cyclic moiety. The cyclic moiety so formedcan be unsubstituted or substituted. Non-limiting exemplary cyclicmoieties includes but are not limited to morpholinyl, azetidinyl,pyrrolidinyl, piperidinyl, piperazinyl, isothiazolidinyl 1,2, dioxide,and thiomorpholinyl. In other examples of the compounds of Formula I,when R⁴ is alkyl, the alkyl is substituted with heterocycloalkyl, whichincludes oxetanyl, azetidinyl, tetrahydrofuranyl, pyrrolyl,tetrahydropyranyl, piperidinyl, morpholinyl, and piperazinyl. All of theabove listed heterocycloaklyl substituents can be unsubstituted orsubstituted.

In yet other examples of the compounds of Formula I, when R⁴ is alkyl,the alkyl is substituted with a 5, 6, 7, 8, 9, or 10 membered monocyclicor bicyclic heteroaryl, which is unsubstituted or substituted. Themonocyclic heteroaryl includes but is not limited to pyrrolyl, thienyl,furyl, pyridinyl, pyranyl, pyrimidinyl, pyrazinyl, pyridazinyl,imidazolyl, thiazolyl, pyrazolyl, and oxazolyl. The bicyclic heteroarylincludes but is not limited benzothiophenyl, benzofuryl, indolyl,quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, benzothiazolyl,quinazolinyl, azaindolyl, pyrazolopyrimidinyl, purinyl, pyrrolo [1,2-b]pyridazinyl, pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl,imidazo[1, 2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

In some embodiments of the compound of Formula I, W² is NR⁴, wherein R⁴is hydrogen, unsubstituted or substituted C₁-C₁₀alkyl (which includesbut is not limited to —CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl,tert-butyl, sec-butyl, pentyl, hexyl, and heptyl), or unsubstituted orsubstituted C₃-C₇cycloalkyl (which includes but is not limited tocyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl). In otherembodiments of the compound of Formula I, R⁴ is unsubstituted orsubstituted heterocycloalkyl (which includes but is not limited tooxetanyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl,piperidinyl, and piperazinyl), or unsubstituted or substitutedC₂-C₁₀heteroalkyl (which includes but is not limited to methoxyethoxy,methoxymethyl, and diethylaminoethyl). Alternatively, R⁴ isunsubstituted or substituted monocyclic heteroaryl (which includes butis not limited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl,pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl,and oxazolyl) or unsubstituted or substituted monocyclic aryl.

In some embodiments R³ and R⁴ taken together form a cyclic moiety. Sucha moiety may have, for example, from 3 to 8 ring atoms. The cyclicmoiety so formed may further include one or more heteroatoms which areselected from the group consisting of S, O, and N. The cyclic moiety soformed is unsubstituted or substituted. In some embodiments, thesubstituent is C₁-C₁₀alkyl (which includes but is not limited to —CH₃,—CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl, pentyl,hexyl, and heptyl), or C₃-C₇cycloalkyl (which includes but is notlimited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl);heterocycloalkyl (which includes but is not limited to oxetanyl,tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperidinyl, andpiperazinyl), C₂-C₁₀heteroalkyl (which includes but is not limited tomethoxyethoxy, methoxymethyl, and diethylaminoethyl); monocyclicheteroaryl (which includes but is not limited to pyrrolyl, thienyl,furyl, pyridinyl, pyranyl, pyrimidinyl, pyrazinyl, pyridazinyl,imidazolyl, thiazolyl, pyrazolyl, and oxazolyl) or unsubstituted orsubstituted monocyclic aryl. The cyclic moiety may have one or moresubstituents, which may be the same or different.

In some embodiments, the cyclic moiety formed by R³ and R⁴ issubstituted with at least one of the following substituents:

In some embodiments of the compound of Formula I, W³ is CR⁵. R⁵ can be,for example, hydrogen, or unsubstituted or substituted alkyl (includingbut not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl,tert-butyl, sec-butyl, pentyl, hexyl, and heptyl). In one embodiment, R⁵is H. In other embodiments, R⁵ is unsubstituted or substituted alkenyl(including but not limited to unsubstituted or substituted C₂-C₅alkenylsuch as, for example, vinyl, allyl, 1-methyl propen-1-yl, butenyl, orpentenyl) or unsubstituted or substituted alkynyl (including but notlimited to unsubstituted or substituted C₁-C₅alkynyl such as acetylenyl,propargyl, butynyl, or pentynyl). Alternatively, R⁵ is unsubstituted orsubstituted aryl (including but not limited to monocyclic or bicyclicaryl) or unsubstituted or substituted arylalkyl (including but notlimited to monocyclic or bicyclic aryl linked to alkyl wherein alkylincludes but is not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl,n-butyl, sec-butyl, and pentyl). In some other embodiments, R⁵ isunsubstituted or substituted heteroaryl, including but not limited tomonocyclic and bicyclic heteroaryl. Monocyclic heteroaryl R⁵ includesbut is not limited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl,pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl,and oxazolyl. Bicyclic heteroaryl R⁵ includes but is not limited tobenzothiophenyl, benzofuryl, indolyl, quinolinyl, isoquinolinyl,benzimidazolyl, benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

In some embodiments of the compound of Formula I, W³ is N or NR⁵,wherein R⁵ is hydrogen, unsubstituted or substituted C₁-C₁₀alkyl (whichincludes but is not limited to —CH₃, —CH₂CH₃, n-propyl, isopropyl,n-butyl, tert-butyl sec-butyl, pentyl, hexyl, and heptyl), orunsubstituted or substituted C₃-C₇cycloalkyl (which includes but is notlimited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl). Inother embodiments of the compound of Formula I, R⁵ is unsubstituted orsubstituted heterocycloalkyl (which includes but is not limited tooxetanyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl,piperidinyl, and piperazinyl), or unsubstituted or substitutedC₂-C₁₀heteroalkyl (which includes but is not limited to methoxyethoxy,methoxymethyl, and diethylaminoethyl). Alternatively, R⁵ isunsubstituted or substituted monocyclic heteroaryl (which includes butis not limited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl,pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl,and oxazolyl) or unsubstituted or substituted monocyclic aryl.

In some embodiments of the compound of Formula I, W⁴ is CR⁶. R⁶ can be,for example, hydrogen, or unsubstituted or substituted alkyl (includingbut not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl,tert-butyl, sec-butyl, pentyl, hexyl, and heptyl). In one embodiment, R⁶is H. In other embodiments, R⁶ is unsubstituted or substituted alkenyl(including but not limited to unsubstituted or substituted C₂-C₅alkenylsuch as, for example, vinyl, allyl, 1-methyl propen-1-yl, butenyl, orpentenyl) or unsubstituted or substituted alkynyl (including but notlimited to unsubstituted or substituted C₂-C₅alkynyl such as acetylenyl,propargyl, butynyl, or pentynyl). Alternatively, R⁶ is unsubstituted orsubstituted aryl (including but not limited to monocyclic or bicyclicaryl) or unsubstituted or substituted arylalkyl (including but notlimited to monocyclic or bicyclic aryl linked to alkyl wherein alkylincludes but is not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl,n-butyl, sec-butyl, and pentyl). In some other embodiments, R⁶ isunsubstituted or substituted heteroaryl, including but not limited tomonocyclic and bicyclic heteroaryl. Monocyclic heteroaryl R⁶ includesbut is not limited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl,pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl,and oxazolyl. Bicyclic heteroaryl R⁶ includes but is not limited tobenzothiophenyl, benzofuryl, indolyl, quinolinyl, isoquinolinyl,benzimidazolyl, benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

In some embodiments of the compound of Formula I, W⁴ is N or NR⁶,wherein R⁶ is hydrogen, unsubstituted or substituted C₁-C₁₀alkyl (whichincludes but is not limited to —CH₃, —CH₂CH₃, n-propyl, isopropyl,n-butyl, tert-butyl, sec-butyl, pentyl, hexyl, and heptyl), orunsubstituted or substituted C₁-C₇cycloalkyl (which includes but is notlimited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl). Inother embodiments of the compound of Formula I, R⁶ is unsubstituted orsubstituted heterocycloalkyl (which includes but is not limited tooxetanyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl,piperidinyl, and piperazinyl), or unsubstituted or substitutedC₂-C₁₀heteroalkyl (which includes but is not limited to methoxyethoxy,methoxymethyl, and diethylaminoethyl). Alternatively, R⁶ isunsubstituted or substituted monocyclic heteroaryl (which includes butis not limited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl,pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl,and oxazolyl) or unsubstituted or substituted monocyclic aryl.

In other embodiments, W⁴ is C═O.

In some embodiments of the compound of Formula I, W⁵ is N. In otherembodiments of the compound of Formula I, W⁵ is CR⁷. R⁷ can be, forexample, hydrogen, or unsubstituted or substituted alkyl (including butnot limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl,sec-butyl, pentyl, hexyl, and heptyl). In one embodiment, R⁷ is H. Inother embodiments, R⁷ is unsubstituted or substituted alkenyl (includingbut not limited to unsubstituted or substituted C₂-C₅alkenyl such as,for example, vinyl, allyl, 1-methyl propen-1-yl, butenyl, or pentenyl)or unsubstituted or substituted alkynyl (including but not limited tounsubstituted or substituted C₂-C₅alkynyl such as acetylenyl, propargyl,butynyl, or pentynyl). Alternatively, R⁷ is unsubstituted or substitutedaryl (including but not limited to monocyclic or bicyclic aryl) orunsubstituted or substituted arylalkyl (including but not limited tomonocyclic or bicyclic aryl linked to alkyl wherein alkyl includes butis not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl,and pentyl). In some other embodiments, R⁷ is unsubstituted orsubstituted heteroaryl, including but not limited to monocyclic andbicyclic heteroaryl. Monocyclic heteroaryl R⁷ includes but is notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl.Bicyclic heteroaryl R⁷ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

In some embodiments of the compound of Formula I, W⁶ is N. In otherembodiments of the compound of Formula I. W is CR⁸. R⁸ can be, forexample, hydrogen, or unsubstituted or substituted alkyl (including butnot limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl,sec-butyl, pentyl, hexyl, and heptyl). In one embodiment, R⁸ is H. Inother embodiments, R⁸ is unsubstituted or substituted alkenyl (includingbut not limited to unsubstituted or substituted C₂-C₅alkenyl such as,for example, vinyl, allyl, 1-methyl propen-1-yl, butenyl, or pentenyl)or unsubstituted or substituted alkynyl (including but not limited tounsubstituted or substituted C₂-C₅alkynyl such as acetylenyl, propargyl,butynyl, or pentynyl). Alternatively, R⁸ is unsubstituted or substitutedaryl (including but not limited to monocyclic or bicyclic aryl) orunsubstituted or substituted arylalkyl (including but not limited tomonocyclic or bicyclic aryl linked to alkyl wherein alkyl includes butis not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl,and pentyl). In some other embodiments, R⁸ is unsubstituted orsubstituted heteroaryl, including but not limited to monocyclic andbicyclic heteroaryl. Monocyclic heteroaryl R⁸ includes but is notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl.Bicyclic heteroaryl R⁸ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

In another aspect, the invention provides compounds of Subformula Ia andIb, where W¹ is CR³, W² is CR⁴, W³ is CR⁴, W⁴ is N, W⁵ is CR⁷, and W⁶ isCR⁸. In one embodiment, R¹, R³, R⁴, R⁵, R⁷ and R⁸ are hydrogen. Inanother embodiment, R¹, R⁴, R⁵, R⁷ and R⁸ are hydrogen and R³ is alkyl,heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy,amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′ and R″are taken together with nitrogen to form a cyclic moiety. R³ can be, forexample, hydrogen, unsubstituted or substituted alkyl (including but notlimited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl,sec-butyl, pentyl, hexyl, and heptyl). In other embodiments, R³ isunsubstituted or substituted alkenyl (including but not limited tounsubstituted or substituted C₂-C₅alkenyl such as, for example, vinyl,allyl, 1-methyl propen-1-yl, butenyl, or pentenyl) or unsubstituted orsubstituted alkynyl (including but not limited to unsubstituted orsubstituted C₂-C₅alkynyl such as acetylenyl, propargyl, butynyl, orpentynyl). Alternatively, R³ is unsubstituted or substituted aryl(including but not limited to monocyclic or bicyclic aryl) orunsubstituted or substituted arylalkyl (including but not limited tomonocyclic or bicyclic aryl linked to alkyl wherein alkyl includes butis not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl,and pentyl). In some other embodiments, R³ is unsubstituted orsubstituted heteroaryl, including but not limited to monocyclic andbicyclic heteroaryl. Monocyclic heteroaryl R³ includes but is notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl.Bicyclic heteroaryl R³ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl. The presentinvention also provides compounds of Formula I wherein R³ isunsubstituted or substituted heteroarylalkyl, including but not limitedto monocyclic and bicyclic heteroaryl as described above, that arelinked to alkyl, which in turn includes but is not limited to CH₃,—CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl. In someembodiments, R³ is unsubstituted or substituted cycloalkyl (includingbut not limited to cyclopropyl, cyclobutyl, and cyclopentyl) orunsubstituted or substituted heteroalkyl (non-limiting examples includeethoxymethyl, methoxymethyl, and diethylaminomethyl). In some furtherembodiments, R³ is unsubstituted or substituted heterocycloalkyl whichincludes but is not limited to pyrrolidinyl, tetrahydrofuranyl,piperidinyl, tetrahydropyranyl, thiazolidinyl, imidazolidinyl,morpholinyl, and piperazinyl. In yet other embodiments of the compoundsof Formula I, R³ is unsubstituted or substituted alkoxy including butnot limited to C₁-C₄alkoxy such as methoxy, ethoxy, propoxy or butoxy.R³ can also be unsubstituted or substituted heterocycloalkyloxy,including but not limited to 4-NH piperidin-1-yl-oxy, 4-methylpiperidin-1-yl-oxy, 4-ethyl piperidin-1-yl-oxy,4-isopropyl-piperidin-1-yl-oxy, and pyrrolidin-3-yl-oxy. In otherembodiments, R³ is unsubstituted or substituted amino, wherein thesubstituted amino includes but is not limited to dimethylamino,diethylamino, di-isopropyl amino, N-methyl N-ethyl amino, anddibutylamino. In some embodiments, R³ is unsubstituted or substitutedacyl, unsubstituted or substituted acyloxy, unsubstituted or substitutedC₁-C₄acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstitutedor substituted amido, or unsubstituted or substituted sulfonamido. Inother embodiments, R³ is halo, which is I, —F, —Cl, or —Br. In someembodiments, R³ is selected from the group consisting of cyano, hydroxy,nitro, phosphate, urea, and carbonate. Also contemplated are R³ being—CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, heptyl, —OCH₃, —OCH₂CH₃, or —CF₃. In some embodiments R³can also be NR′R″ wherein R′ and R″ are taken together with the nitrogento form a cyclic moiety having from 3 to 8 ring atoms. The cyclic moietyso formed may further include one or more heteroatoms which are selectedfrom the group consisting of S, O, and N. The cyclic moiety so formed isunsubstituted or substituted, including but not limited to morpholinyl,azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, isothiazolidinyl1,2, dioxide, and thiomorpholinyl. Further non-limiting exemplary cyclicmoieites are the following:

In another aspect, the invention provides compounds of Subformula Ic andId, where W¹ is N, W² is CR⁴, W³ is CR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ isCR⁸. In one embodiment, R¹, R⁴, R⁵, R⁷ and R⁸ are hydrogen. In anotherembodiment, R¹, R⁵, R⁷ and R⁸ are hydrogen and R⁴ is alkyl, heteroalkyl,alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are takentogether with nitrogen to form a cyclic moiety. R⁴ can be, for example,hydrogen, unsubstituted or substituted alkyl (including but not limitedto CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, and heptyl). In other embodiments, R⁴ is unsubstituted orsubstituted alkenyl (including but not limited to unsubstituted orsubstituted C₂-C₅alkenyl such as, for example, vinyl, allyl, 1-methylpropen-1-yl, butenyl, or pentenyl) or unsubstituted or substitutedalkynyl (including but not limited to unsubstituted or substitutedC₂-C₅alkynyl such as acetylenyl, propargyl, butynyl, or pentynyl).Alternatively, R⁴ is unsubstituted or substituted aryl (including butnot limited to monocyclic or bicyclic aryl) or unsubstituted orsubstituted arylalkyl (including but not limited to monocyclic orbicyclic aryl linked to alkyl wherein alkyl includes but is not limitedto CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl).In some other embodiments, R⁴ is unsubstituted or substitutedheteroaryl, including but not limited to monocyclic and bicyclicheteroaryl. Monocyclic heteroaryl R⁴ includes but is not limited topyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl, pyrazinyl,pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl. Bicyclicheteroaryl R⁴ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl. The presentinvention also provides compounds of Formula I wherein R⁴ isunsubstituted or substituted heteroarylalkyl, including but not limitedto monocyclic and bicyclic heteroaryl as described above, that arelinked to alkyl, which in turn includes but is not limited to CH₃,—CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl. In someembodiments, R⁴ is unsubstituted or substituted cycloalkyl (includingbut not limited to cyclopropyl, cyclobutyl, and cyclopentyl) orunsubstituted or substituted heteroalkyl (non-limiting examples includeethoxymethyl, methoxymethyl, and diethylaminomethyl). In some furtherembodiments, R⁴ is unsubstituted or substituted heterocycloalkyl whichincludes but is not limited to pyrrolidinyl, tetrahydrofuranyl,piperidinyl, tetrahydropyranyl, thiazolidinyl, imidazolidinyl,morpholinyl, and piperazinyl. In yet other embodiments of the compoundsof Formula I, R⁴ is unsubstituted or substituted alkoxy including butnot limited to C₁-C₄alkoxy such as methoxy, ethoxy, propoxy or butoxy.R³ can also be unsubstituted or substituted heterocycloalkyloxy,including but not limited to 4-NH piperidin-1-yl-oxy, 4-methylpiperidin-1-yl-oxy, 4-ethyl piperidin-1-yl-oxy,4-isopropyl-piperidin-1-yl-oxy, and pyrrolidin-3-yl-oxy. In otherembodiments, R⁴ is unsubstituted or substituted amino, wherein thesubstituted amino includes but is not limited to dimethylamino,diethylamino, di-isopropyl amino, N-methyl N-ethyl amino, anddibutylamino. In some embodiments, R⁴ is unsubstituted or substitutedacyl, unsubstituted or substituted acyloxy, unsubstituted or substitutedC₁-C₄acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstitutedor substituted amido, or unsubstituted or substituted sulfonamido. Inother embodiments, R⁴ is halo, which is —I, —F, —Cl, or —Br. In someembodiments, R⁴ is selected from the group consisting of cyano, hydroxy,nitro, phosphate, urea, and carbonate. Also contemplated are R⁴ being—CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, heptyl, —OCH₃, —OCH₂CH₃, or —CF₃. In some embodiments R⁴can also be NR′R″ wherein R′ and R″ are taken together with the nitrogento form a cyclic moiety having from 3 to 8 ring atoms. The cyclic moietyso formed may further include one or more heteroatoms which are selectedfrom the group consisting of S, O, and N. The cyclic moiety so formed isunsubstituted or substituted, including but not limited to morpholinyl,azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, isothiazolidinyl1,2, dioxide, and thiomorpholinyl. Further non-limiting exemplary cyclicmoieites are the following:

In another aspect, the invention provides compounds of Subformula Ie andIf, where W¹ is CR³, W² is N, W³ is CR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ isCR⁸.

In one embodiment, R¹, R³, R⁵, R⁷ and R⁸ are hydrogen. In anotherembodiment, R¹, R⁵, R⁷ and R⁸ are hydrogen and R³ is alkyl, heteroalkyl,alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are takentogether with nitrogen to form a cyclic moiety. R³ can be, for example,hydrogen, unsubstituted or substituted alkyl (including but not limitedto CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, and heptyl). In other embodiments, R³ is unsubstituted orsubstituted alkenyl (including but not limited to unsubstituted orsubstituted C₂-C₅alkenyl such as, for example, vinyl, allyl, 1-methylpropen-1-yl, butenyl, or pentenyl) or unsubstituted or substitutedalkynyl (including but not limited to unsubstituted or substitutedC₂-C₅alkynyl such as acetylenyl, propargyl, butynyl, or pentynyl).Alternatively, R³ is unsubstituted or substituted aryl (including butnot limited to monocyclic or bicyclic aryl) or unsubstituted orsubstituted arylalkyl (including but not limited to monocyclic orbicyclic aryl linked to alkyl wherein alkyl includes but is not limitedto CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl).In some other embodiments, R³ is unsubstituted or substitutedheteroaryl, including but not limited to monocyclic and bicyclicheteroaryl. Monocyclic heteroaryl R³ includes but is not limited topyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl, pyrazinyl,pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl. Bicyclicheteroaryl R³ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl. The presentinvention also provides compounds of Formula I wherein R³ isunsubstituted or substituted heteroarylalkyl, including but not limitedto monocyclic and bicyclic heteroaryl as described above, that arelinked to alkyl, which in turn includes but is not limited to CH₃,—CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl. In someembodiments, R³ is unsubstituted or substituted cycloalkyl (includingbut not limited to cyclopropyl, cyclobutyl, and cyclopentyl) orunsubstituted or substituted heteroalkyl (non-limiting examples includeethoxymethyl, methoxymethyl, and diethylaminomethyl). In some furtherembodiments, R³ is unsubstituted or substituted heterocycloalkyl whichincludes but is not limited to pyrrolidinyl, tetrahydrofuranyl,piperidinyl, tetrahydropyranyl, thiazolidinyl, imidazolidinyl,morpholinyl, and piperazinyl. In yet other embodiments of the compoundsof Formula I, R³ is unsubstituted or substituted alkoxy including butnot limited to C₁-C₄alkoxy such as methoxy, ethoxy, propoxy or butoxy.R³ can also be unsubstituted or substituted heterocycloalkyloxy,including but not limited to 4-NH piperidin-1-yl-oxy, 4-methylpiperidin-1-yl-oxy, 4-ethyl piperidin-1-yl-oxy,4-isopropyl-piperidin-1-yl-oxy, and pyrrolidin-3-yl-oxy. In otherembodiments, R³ is unsubstituted or substituted amino, wherein thesubstituted amino includes but is not limited to dimethylamino,diethylamino, di-isopropyl amino, N-methyl N-ethyl amino, anddibutylamino. In some embodiments, R³ is unsubstituted or substitutedacyl, unsubstituted or substituted acyloxy, unsubstituted or substitutedC₁-C₄acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstitutedor substituted amido, or unsubstituted or substituted sulfonamido. Inother embodiments, R³ is halo, which is —I, —F, —Cl, or —Br. In someembodiments, R³ is selected from the group consisting of cyano, hydroxy,nitro, phosphate, urea, and carbonate. Also contemplated are R³ being—CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, heptyl, —OCH₃, —OCH₂CH₃, or —CF₃. In some embodiments R³can also be NR′R″ wherein R′ and R″ are taken together with the nitrogento form a cyclic moiety having from 3 to 8 ring atoms. The cyclic moietyso formed may further include one or more heteroatoms which are selectedfrom the group consisting of S, O, and N. The cyclic moiety so formed isunsubstituted or substituted, including but not limited to morpholinyl,azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, isothiazolidinyl1,2, dioxide, and thiomorpholinyl. Further non-limiting exemplary cyclicmoieites are the following:

In some embodiments, the substituents R³, R⁴, R⁵, or R⁶ may be any ofthe substituents shown in Table 1:

TABLE 1 R³, R⁴, R⁵, R⁶ moieties of the compounds of Formula I, eachindependently includes but is not limited to the following: Sub- class #R R-1 

R-2 

R-3 

R-4 

R-5 

R-6 

R-7 

R-8 

R-9  —CH(CH₃)₂ R-10

R-11

R-12

R-13

R-14

R-15

R-16

R-17

R-18

R-19

R-20

R-21

R-22

R-23

R-24

R-25

R-26

R-27

R-28

R-29

R-30

R-31

R-32

R-33

R-34

R-35

R-36

R-37

R-38

R-39

R-40 H R-41

R-42

R-43

R-44

R-45

R-46

R-47

R-48

R-49

R-50

R-51

R-52

R-53

R-54

R-55

R-56

R-57

R-58

R-59

R-60

R-61

R-62

R-63

R-64

R-65

R-66

R-67

R-68

R-69

R-70

R-71

R-72

R-73

R-74

R-75

R-76

R-77

R-78

R-79

R-80

R-81

In another aspect, the invention provides a compound of Formula II:

or its pharmaceutically acceptable salts thereof, where:

-   -   X is O or S or N;    -   W¹ is S, N, NR³ or CR³, W² is N or CR⁴, W³ is S, N or CR⁵, W⁴ is        N or C, and W⁷ is N or C, wherein no more than two N atoms and        no more than two C═O groups are adjacent;    -   W⁵ is N or CR⁷;    -   W⁶ is N or CR⁸;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   or R³ and R⁴ taken together form a cyclic moiety; and    -   R⁵, R⁷ and R⁸ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

In some embodiments, the compound of Formula II exists as a tautomer,and such tautomers are contemplated by the present invention.

In some embodiments, the compound of Formula II has the Formula:

In yet other embodiments, W¹ is CR³, W² is CR⁴, W³ is N, W⁴ is N, W⁵ isCR⁷, and W⁶ is CR⁸. In other embodiments, W¹ is CR³, W² is CR⁴, W³ is N,W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸. In other embodiments, W¹ is CR³, W²is CR⁴, W³ is N, W⁴ is N, W⁵ is N, and W⁶ is CR⁸. In still otherembodiments, W¹ is NR³, W² is CR⁴, W³ is N, W⁴ is C, W⁵ is CR⁷, and W⁶is CR⁸. In other embodiments, W¹ is S, W² is CR⁴, W³ is N, W⁴ is C, W⁵is CR⁷, and W⁶ is CR^(X). In other embodiments, W¹ is CR⁴, W² is CR⁴, W³is S, W⁴ is C, W⁶ is N, and W⁶ is N.

In other embodiments, a compound of Formula II is a compound accordingto one of the formulas:

In some embodiments, X is O. In other embodiments, X is S.

In some embodiments, R¹ is hydrogen. In other embodiments, R¹ is alkyl,heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy,amido, amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,hydroxy, nitro, phosphate, urea, carbonate, or NR′R″, wherein R′ and R″are taken together with nitrogen to form a cyclic moiety.

In some embodiments, R² is hydrogen. In other embodiments, R² is, forexample, unsubstituted or substituted alkyl (including but not limitedto CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butylpentyl, hexyl, and heptyl). In other embodiments, R² is unsubstituted orsubstituted alkenyl (including but not limited to unsubstituted orsubstituted C₂-C₅alkenyl such as, for example, vinyl, allyl, 1-methylpropen-1-yl, butenyl, or pentenyl) or unsubstituted or substitutedalkynyl (including but not limited to unsubstituted or substitutedC₂-C₅alkynyl such as acetylenyl, propargyl, butynyl, or pentynyl).Alternatively, R² is unsubstituted or substituted aryl (including butnot limited to monocyclic or bicyclic aryl) or unsubstituted orsubstituted arylalkyl (including but not limited to monocyclic orbicyclic aryl linked to alkyl wherein alkyl includes but is not limitedto CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl).In some other embodiments, R² is unsubstituted or substitutedheteroaryl, including but not limited to monocyclic and bicyclicheteroaryl. Monocyclic heteroaryl R² includes but is not limited topyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl, pyrazinyl,pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl. Bicyclicheteroaryl R² includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl. The presentinvention also provides compounds wherein R² is unsubstituted orsubstituted heteroarylalkyl, including but not limited to monocyclic andbicyclic heteroaryl as described above, that are linked to alkyl, whichin turn includes but is not limited to CH₃, —CH₂CH₃, n-propyl,isopropyl, n-butyl, sec-butyl, and pentyl. In some embodiments, R² isunsubstituted or substituted cycloalkyl (including but not limited tocyclopropyl, cyclobutyl, and cyclopentyl) or unsubstituted orsubstituted heteroalkyl (non-limiting examples include ethoxymethyl,methoxymethyl, and diethylaminomethyl). In some further embodiments, R²is unsubstituted or substituted heterocycloalkyl which includes but isnot limited to pyrrolidinyl, tetrahydrofuranyl, piperidinyl,tetrahydropyranyl, thiazolidinyl, imidazolidinyl, morpholinyl, andpiperazinyl. In yet other embodiments of the compounds of Formula II, R²is unsubstituted or substituted alkoxy including but not limited toC₁-C₄alkoxy such as methoxy, ethoxy, propoxy or butoxy. R² can also beunsubstituted or substituted heterocycloalkyloxy, including but notlimited to 4-NH piperidin-1-yl-oxy, 4-methyl piperidin-1-yl-oxy, 4-ethylpiperidin-1-yl-oxy, 4-isopropyl-piperidin-1-yl-oxy, andpyrrolidin-3-yl-oxy. In other embodiments, R² is unsubstituted orsubstituted amino, wherein the substituted amino includes but is notlimited to dimethylamino, diethylamino, di-isopropyl amino, N-methylN-ethyl amino, and dibutylamino. In some embodiments, R² isunsubstituted or substituted acyl, unsubstituted or substituted acyloxy,unsubstituted or substituted C₁-C₄acyloxy, unsubstituted or substitutedalkoxycarbonyl, unsubstituted or substituted amido, or unsubstituted orsubstituted sulfonamido. In other embodiments, R² is halo, which is —I,—F, —Cl, or —Br. In some embodiments, R² is selected from the groupconsisting of cyano, hydroxy, nitro, phosphate, urea, and carbonate.Also contemplated are R² being —CH₃, —CH₂CH₃, n-propyl, isopropyl,n-butyl, tert-butyl, sec-butyl, pentyl, hexyl, heptyl, —OCH₃, —OCH₂CH₃,or —CF₃.

In some embodiments of the compound of Formula II, W¹ is CR³. R³ can be,for example, hydrogen, unsubstituted or substituted alkyl (including butnot limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl,sec-butyl, pentyl, hexyl, and heptyl). In other embodiments, R³ isunsubstituted or substituted alkenyl (including but not limited tounsubstituted or substituted C₂-C₅alkenyl such as, for example, vinyl,allyl, 1-methyl propen-1-yl, butenyl, or pentenyl) or unsubstituted orsubstituted alkynyl (including but not limited to unsubstituted orsubstituted C₂-C₅alkynyl such as acetylenyl, propargyl, butynyl, orpentynyl). Alternatively, R³ is unsubstituted or substituted aryl(including but not limited to monocyclic or bicyclic aryl) orunsubstituted or substituted arylalkyl (including but not limited tomonocyclic or bicyclic aryl linked to alkyl wherein alkyl includes butis not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl,and pentyl). In some other embodiments, R³ is unsubstituted orsubstituted heteroaryl, including but not limited to monocyclic andbicyclic heteroaryl. Monocyclic heteroaryl R³ includes but is notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl.Bicyclic heteroaryl R³ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl. The presentinvention also provides compounds of Formula II wherein R³ isunsubstituted or substituted heteroarylalkyl, including but not limitedto monocyclic and bicyclic heteroaryl as described above, that arelinked to alkyl which in turn includes but is not limited to CH₃,—CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl. In someembodiments, R³ is unsubstituted or substituted cycloalkyl (includingbut not limited to cyclopropyl, cyclobutyl, and cyclopentyl) orunsubstituted or substituted heteroalkyl (non-limiting examples includeethoxymethyl, methoxymethyl, and diethylaminomethyl). In some furtherembodiments, R³ is unsubstituted or substituted heterocycloalkyl whichincludes but is not limited to pyrrolidinyl, tetrahydrofuranyl,piperidinyl, tetrahydropyranyl, thiazolidinyl, imidazolidinyl,morpholinyl, and piperazinyl. In yet other embodiments of the compoundsof Formula II, R³ is unsubstituted or substituted alkoxy including butnot limited to C₁-C₄alkoxy such as methoxy, ethoxy, propoxy or butoxy.R³ can also be unsubstituted or substituted heterocycloalkyloxy,including but not limited to 4-NH piperidin-1-yl-oxy, 4-methylpiperidin-1-yl-oxy, 4-ethyl piperidin-1-yl-oxy,4-isopropyl-piperidin-1-yl-oxy, and pyrrolidin-3-yl-oxy. In otherembodiments, R³ is unsubstituted or substituted amino, wherein thesubstituted amino includes but is not limited to dimethylamino,diethylamino, di-isopropyl amino, N-methyl N-ethyl amino, anddibutylamino. In some embodiments, R³ is unsubstituted or substitutedacyl, unsubstituted or substituted acyloxy, unsubstituted or substitutedC₁-C₄acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstitutedor substituted amido, or unsubstituted or substituted sulfonamido. Inother embodiments, R³ is halo, which is —I, —F, —Cl, or —Br. In someembodiments, R³ is selected from the group consisting of cyano, hydroxy,nitro, phosphate, urea, and carbonate. Also contemplated are R³ being—CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, heptyl, —OCH₃, —OCH₂CH, or —CF₃.

R³ of the compounds of Formula II, can also be NR′R″ wherein R′ and R″are taken together with the nitrogen to form a cyclic moiety having from3 to 8 ring atoms. The cyclic moiety so formed may further include oneor more heteroatoms which are selected from the group consisting of S,O, and N. The cyclic moiety so formed is unsubstituted or substituted,including but not limited to morpholinyl, azetidinyl, pyrrolidinyl,piperidinyl, piperazinyl, isothiazolidinyl 1,2, dioxide, andthiomorpholinyl. Further non-limiting exemplary cyclic moieites are thefollowing:

The invention also provides compounds of Formula II, wherein when R³ isa member of the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl,heteroalkyl, heterocycloalkyl, heterocycloalkyloxy, aryl, arylalkyl,heteroaryl, heteroarylalkyl, acyl, alkoxy, amido, amino, sulfonamido,acyloxy, alkoxycarbonyl, and NR′R″ (wherein R′ and R″ are taken togetherwith nitrogen to form a cyclic moiety), then R³ is optionallysubstituted with one or more of the following substituents: alkyl,alkenyl, alkynyl, cycloalkyl, heteroalkyl, heterocycloalkyl,heterocycloalkyloxy, aryl, arylalkyl, heteroaryl, heteroarylalkyl, acyl,heterocycloalkyloxy, alkoxy, amido, amino, sulfonamido, acyloxy,alkoxycarbonyl, halo, cyano, hydroxy, nitro, phosphate, urea, carbonate,or NR′R″ wherein R′ and R″ are taken together with nitrogen to form acyclic moiety. Each of the above substituents may be further substitutedwith one or more substituents chosen from the group consisting of alkyl,alkoxy, amido, amino, sulfonamido, acyloxy, alkoxycarbonyl, halo, cyano,hydroxy, nitro, oxo, phosphate, urea, and carbonate.

For example, the invention provides compounds wherein when R³ is alkyl,the alkyl is substituted with NR′R″ wherein R′ and R″ are taken togetherwith the nitrogen to form a cyclic moiety. The cyclic moiety so formedcan be unsubstituted or substituted. Non-limiting exemplary cyclicmoieties includes but are not limited to morpholinyl, azetidinyl,pyrrolidinyl, piperidinyl, piperazinyl, and thiomorpholinyl. In otherexamples of the compounds of Formula II, when R³ is alkyl, the alkyl issubstituted with heterocycloalkyl, which includes oxetanyl, azetidinyl,tetrahydrofuranyl, pyrrolyl, tetrahydropyranyl, piperidinyl,morpholinyl, and piperazinyl. All of the above listed heterocycloaklylsubstituents can be unsubstituted or substituted.

In yet other examples of the compounds of Formula II, when R³ is alkyl,the alkyl is substituted with a 5, 6, 7, 8, 9, or 10 membered monocyclicor bicyclic heteroaryl, which is unsubstituted or substituted. Themonocyclic heteroaryl includes but is not limited to pyrrolyl, thienyl,furyl, pyridinyl, pyranyl, pyrimidinyl, pyrazinyl, pyridazinyl,imidazolyl, thiazolyl, pyrazolyl, and oxazolyl. The bicyclic heteroarylincludes but is not limited to benzothiophenyl, benzofuryl, indolyl,quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, benzothiazolyl,quinazolinyl, azaindolyl, pyrazolopyrimidinyl, purinyl, pyrrolo [1,2-b]pyridazinyl, pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl,imidazo[1, 2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

In other embodiments of the compound of Formula II, R³ is —NHR^(3′),—N(CH₃)R^(3′)—N(CH₂CH₃)R^(3′), —N(CH(CH₃)₂)R^(3′), or —OR^(3′), whereinR^(3′) is unsubstituted or substituted heterocycloalkyl (nonlimitingexamples thereof include 4-NH piperidin-1-yl, 4-methyl piperidin-1-yl,4-ethyl piperidin-1-yl, 4-isopropyl-piperidin-1-yl, andpyrrolidin-3-yl), unsubstituted or substituted monocyclic aryl, orunsubstituted or substituted monocyclic heteroaryl (including but notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl).In one example, R³ is O-aryl, i.e. phenoxy. In another example, R³ is—O-(4-methyl)piperidin-1-yl or —O-(4-isopropyl)piperidin-1-yl.

In some embodiments of the compound of Formula II, R³ is one of thefollowing moieties:

In some embodiments of the compound of Formula II, W¹ is NR³, wherein R³is hydrogen, unsubstituted or substituted C₁-C₁₀alkyl (which includesbut is not limited to —CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyltert-butyl, sec-butyl, pentyl, hexyl, and heptyl), or unsubstituted orsubstituted C₃-C₇cycloalkyl (which includes but is not limited tocyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl). In otherembodiments of the compound of Formula II, R³ is unsubstituted orsubstituted heterocycloalkyl (which includes but is not limited tooxetanyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl,piperidinyl, and piperazinyl), or unsubstituted or substitutedC₂-C₁₀heteroalkyl (which includes but is not limited to methoxyethoxy,methoxymethyl, and diethylaminoethyl). Alternatively, R³ isunsubstituted or substituted monocyclic heteroaryl (which includes butis not limited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl,pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl,and oxazolyl) or unsubstituted or substituted monocyclic aryl.

In other embodiments, W¹ is N. In still other embodiments, W¹ is S.

In some embodiments of the compound of Formula II. W² is CR⁴. R⁴ can be,for example, hydrogen, or unsubstituted or substituted alkyl (includingbut not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl,tert-butyl, sec-butyl, pentyl, hexyl, and heptyl). In other embodiments,R⁴ is unsubstituted or substituted alkenyl (including but not limited tounsubstituted or substituted C₂-C₅alkenyl such as, for example, vinyl,allyl, 1-methyl propen-1-yl, butenyl, or pentenyl) or unsubstituted orsubstituted alkynyl (including but not limited to unsubstituted orsubstituted C₂-C₅alkynyl such as acetylenyl, propargyl, butynyl, orpentynyl). Alternatively, R⁴ is unsubstituted or substituted aryl(including but not limited to monocyclic or bicyclic aryl) orunsubstituted or substituted arylalkyl (including but not limited tomonocyclic or bicyclic aryl linked to alkyl wherein alkyl includes butis not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl,and pentyl). In some other embodiments, R⁴ is unsubstituted orsubstituted heteroaryl, including but not limited to monocyclic andbicyclic heteroaryl. Monocyclic heteroaryl R⁴ includes but is notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl.Bicyclic heteroaryl R⁴ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

The present invention also provides compounds of Formula II wherein R⁴is unsubstituted or substituted heteroarylalkyl, including but notlimited to monocyclic and bicyclic heteroaryl as described above, thatare linked to alkyl, which in turn includes but is not limited to CH₃,—CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl. In someembodiments, R⁴ is unsubstituted or substituted cycloalkyl (includingbut not limited to cyclopropyl, cyclobutyl, and cyclopentyl) orunsubstituted or substituted heteroalkyl (non-limiting examples includeethoxymethyl, methoxymethyl, and diethylaminomethyl). In some furtherembodiments, R⁴ is unsubstituted or substituted heterocycloalkyl whichincludes but is not limited to pyrrolidinyl, tetrahydrofuranyl,piperidinyl, tetrahydropyranyl, thiazolidinyl, imidazolidinyl,morpholinyl, and piperazinyl. In yet other embodiments of the compoundsof Formula II, R⁴ is unsubstituted or substituted alkoxy including butnot limited to C₁-C₄alkoxy such as methoxy, ethoxy, propoxy or butoxy.R⁴ can also be unsubstituted or substituted heterocycloalkyloxy,including but not limited to 4-NH piperidin-1-yl-oxy, 4-methylpiperidin-1-yl-oxy, 4-ethyl piperidin-1-yl-oxy,4-isopropyl-piperidin-1-yl-oxy, and pyrrolidin-3-yl-oxy. In otherembodiments, R⁴ is unsubstituted or substituted amino, wherein thesubstituted amino includes but is not limited to dimethylamino,diethylamino, di-isopropyl amino, N-methyl N-ethyl amino, anddibutylamino. In some embodiments, R⁴ is unsubstituted or substitutedacyl, unsubstituted or substituted acyloxy, unsubstituted or substitutedC₁-C₄acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstitutedor substituted amido, or unsubstituted or substituted sulfonamido. Insome embodiments, R⁴ is halo, which is —I, —F, —Cl, or —Br. In someembodiments, R⁴ is selected from the group consisting of cyano, hydroxy,nitro, phosphate, urea, or carbonate. Also contemplated are R⁴ being—CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, heptyl, —OCH₃, —OCH₂CH₃, or —CF₃.

R⁴ of the compounds of Formula II, can also be NR′R″ wherein R′ and R″are taken together with the nitrogen to form a cyclic moiety having from3 to 8 ring atoms. The cyclic moiety so formed may further include oneor more heteroatoms which are selected from the group consisting of S,O, and N. The cyclic moiety so formed is unsubstituted or substituted,including but not limited to morpholinyl, azetidinyl, pyrrolidinyl,piperidinyl, piperazinyl, isothiazolidinyl 1,2, dioxide, andthiomorpholinyl. Further non-limiting exemplary cyclic moieties are thefollowing:

The invention also provides compounds of Formula II, wherein when R⁴ isa member of the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl,heteroalkyl, heterocycloalkyl, heterocycloalkyloxy, aryl, arylalkyl,heteroaryl, heteroarylalkyl, acyl, alkoxy, amido, amino, sulfonamido,acyloxy, alkoxycarbonyl, and NR′R″ (wherein R′ and R″ are taken togetherwith nitrogen to form a cyclic moiety), then R⁴ is optionallysubstituted with one or more of the following substituents: alkyl,alkenyl, alkynyl, cycloalkyl, heteroalkyl, heterocycloalkyl,heterocycloalkyloxy, aryl, arylalkyl, heteroaryl, heteroarylalkyl, acyl,alkoxy, amido, amino, sulfonamido, acyloxy, alkoxycarbonyl, halo, cyano,hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′ and R″are taken together with nitrogen to form a cyclic moiety. Each of theabove substituents may be further substituted with one or moresubstituents chosen from the group consisting of alkyl, alkoxy, amido,amino, sulfonamido, acyloxy, alkoxycarbonyl, halo, cyano, hydroxy,nitro, oxo, phosphate, urea, and carbonate.

For example, the invention provides compounds wherein when R⁴ is alkyl,the alkyl is substituted with NR′R″ wherein R′ and R″ are taken togetherwith the nitrogen to form a cyclic moiety. The cyclic moiety so formedcan be unsubstituted or substituted. Non-limiting exemplary cyclicmoieties includes but are not limited to morpholinyl, azetidinyl,pyrrolidinyl, piperidinyl, piperazinyl, isothiazolidinyl 1, 2, dioxide,and thiomorpholinyl. In other examples of the compounds of Formula II,when R⁴ is alkyl, the alkyl is substituted with heterocycloalkyl, whichincludes oxetanyl, azetidinyl, tetrahydrofuranyl, pyrrolyl,tetrahydropyranyl, piperidinyl, morpholinyl, and piperazinyl. All of theabove listed heterocycloaklyl substituents can be unsubstituted orsubstituted.

In yet other examples of the compounds of Formula II, when R⁴ is alkyl,the alkyl is substituted with a 5, 6, 7, 8, 9, or 10 membered monocyclicor bicyclic heteroaryl, which is unsubstituted or substituted. Themonocyclic heteroaryl includes but is not limited to pyrrolyl, thienyl,furyl, pyridinyl, pyranyl, pyrimidinyl, pyrazinyl, pyridazinyl,imidazolyl, thiazolyl, pyrazolyl, and oxazolyl. The bicyclic heteroarylincludes but is not limited benzothiophenyl, benzofuryl, indolyl,quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, benzothiazolyl,quinazolinyl, azaindolyl, pyrazolopyrimidinyl, purinyl, pyrrolo [1,2-b]pyridazinyl, pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl,imidazo[1, 2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl. In someembodiments of the compound of Formula II, W² is N.

In some embodiments R³ and R⁴ taken together form a cyclic moiety. Sucha moiety may have, for example, from 3 to 8 ring atoms. The cyclicmoiety so formed may further include one or more heteroatoms which areselected from the group consisting of S, O, and N. The cyclic moiety soformed is unsubstituted or substituted. In some embodiments, thesubstituent is C₁-C₁₀alkyl (which includes but is not limited to —CH₃,—CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl, pentyl,hexyl, and heptyl), or C₃-C₇cycloalkyl (which includes but is notlimited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl);heterocycloalkyl (which includes but is not limited to oxetanyl,tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperidinyl, andpiperazinyl), C₂-C₁₀heteroalkyl (which includes but is not limited tomethoxyethoxy, methoxymethyl, and diethylaminoethyl); monocyclicheteroaryl (which includes but is not limited to pyrrolyl, thienyl,furyl, pyridinyl, pyranyl, pyrimidinyl, pyrazinyl, pyridazinyl,imidazolyl, thiazolyl, pyrazolyl, and oxazolyl) or unsubstituted orsubstituted monocyclic aryl. The cyclic moiety may have one or moresubstituents, which may be the same or different.

In some embodiments, the cyclic moiety formed by R³ and R⁴ issubstituted with at least one of the following substituents:

In some embodiments of the compound of Formula II, W³ is CR⁵. R⁵ can be,for example, hydrogen, or unsubstituted or substituted alkyl (includingbut not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl,tert-butyl, sec-butyl, pentyl hexyl, and heptyl). In one embodiment, R⁵is H. In other embodiments, R⁵ is unsubstituted or substituted alkenyl(including but not limited to unsubstituted or substituted C₂-C₅alkenylsuch as, for example, vinyl, allyl, 1-methyl propen-1-yl, butenyl, orpentenyl) or unsubstituted or substituted alkynyl (including but notlimited to unsubstituted or substituted C₂-C₅alkynyl such as acetylenyl,propargyl, butynyl, or pentynyl). Alternatively, R⁵ is unsubstituted orsubstituted aryl (including but not limited to monocyclic or bicyclicaryl) or unsubstituted or substituted arylalkyl (including but notlimited to monocyclic or bicyclic aryl linked to alkyl wherein alkylincludes but is not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl,n-butyl, sec-butyl, and pentyl). In some other embodiments, R⁵ isunsubstituted or substituted heteroaryl, including but not limited tomonocyclic and bicyclic heteroaryl. Monocyclic heteroaryl R⁵ includesbut is not limited to pyrrolyl, thienyl, furyl, pyridinyl, pyranylpyrimidinyl, pyrazinyl, pyridazinyl imidazolyl, thiazolyl pyrazolyl, andoxazolyl. Bicyclic heteroaryl R⁵ includes but is not limited tobenzothiophenyl benzofuryl indolyl, quinolinyl isoquinolinyl,benzimidazolyl, benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl. In someembodiments of the compound of Formula II, W³ is N. In otherembodiments, W³ is S.

In some embodiments of the compound of Formula II, W⁴ is C. In otherembodiments, W⁴ is N.

In some embodiments of the compound of Formula II, W⁵ is N. In otherembodiments of the compound of Formula II, W⁵ is CR⁷. R⁷ can be, forexample, hydrogen, or unsubstituted or substituted alkyl (including butnot limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butylsec-butyl, pentyl, hexyl, and heptyl). In one embodiment, R⁷ is H. Inother embodiments, R⁷ is unsubstituted or substituted alkenyl (includingbut not limited to unsubstituted or substituted C₂-C₅alkenyl such as,for example, vinyl, allyl, 1-methyl propen-1-yl, butenyl, or pentenyl)or unsubstituted or substituted alkynyl (including but not limited tounsubstituted or substituted C₂-C₅alkynyl such as acetylenyl, propargyl,butynyl, or pentynyl). Alternatively, R⁷ is unsubstituted or substitutedaryl (including but not limited to monocyclic or bicyclic aryl) orunsubstituted or substituted arylalkyl (including but not limited tomonocyclic or bicyclic aryl linked to alkyl wherein alkyl includes butis not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl,and pentyl). In some other embodiments, R⁷ is unsubstituted orsubstituted heteroaryl, including but not limited to monocyclic andbicyclic heteroaryl. Monocyclic heteroaryl R⁷ includes but is notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl.Bicyclic heteroaryl R⁷ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

In some embodiments of the compound of Formula II, W⁶ is N. In otherembodiments of the compound of Formula II, W⁶ is CR⁸. R⁸ can be, forexample, hydrogen, or unsubstituted or substituted alkyl (including butnot limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl,sec-butyl, pentyl, hexyl, and heptyl). In one embodiment, R⁸ is H. Inother embodiments, R⁸ is unsubstituted or substituted alkenyl (includingbut not limited to unsubstituted or substituted C₂-C₅alkenyl such as,for example, vinyl, allyl, 1-methyl propen-1-yl, butenyl, or pentenyl)or unsubstituted or substituted alkynyl (including but not limited tounsubstituted or substituted C₂-C₅alkynyl such as acetylenyl, propargyl,butynyl, or pentynyl). Alternatively, R⁸ is unsubstituted or substitutedaryl (including but not limited to monocyclic or bicyclic aryl) orunsubstituted or substituted arylalkyl (including but not limited tomonocyclic or bicyclic aryl linked to alkyl wherein alkyl includes butis not limited to CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl,and pentyl). In some other embodiments, R⁸ is unsubstituted orsubstituted heteroaryl, including but not limited to monocyclic andbicyclic heteroaryl. Monocyclic heteroaryl R⁸ includes but is notlimited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl, pyrimidinyl,pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl, and oxazolyl.Bicyclic heteroaryl R⁸ includes but is not limited to benzothiophenyl,benzofuryl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-f][1, 2, 4]triazinyl.

In some embodiments of the compound of Formula II, W⁷ is C. In otherembodiments, W⁷ is N.

The invention also provides compounds of Formula II which are defined asdefined by the following subclasses.

In some embodiments of compounds of Subclasses IIa-IIj, R¹ is hydrogen.In other embodiments of compounds of Subclasses IIa-IIl, R² is NH₂ ofNHCO(alkyl). In other embodiments of compounds of Subclasses IIa-IIl, R⁴is hydrogen. In other embodiments of compounds of Subclasses IIc-IIf andIIi-IIl, R⁷ is hydrogen. In other embodiments of compounds of SubclassesIIa-IIh and IIk-IIl. R⁸ is hydrogen.

In some embodiments of compounds of Subclasses IIa through IIl, R³ isalkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,heterocycloalkyloxy, amido, amino, acyl, acyloxy, alkoxycarbonyl,sulfonamido, halo, cyano, hydroxy, nitro, phosphate, urea, carbonate, orNR′R″ wherein R′ and R″ are taken together with nitrogen to form acyclic moiety. R³ can be, for example, hydrogen, unsubstituted orsubstituted alkyl (including but not limited to CH₃, —CH₂CH₃, n-propyl,isopropyl, n-butyl, tert-butyl, sec-butyl, pentyl, hexyl, and heptyl).In other embodiments, R³ is unsubstituted or substituted alkenyl(including but not limited to unsubstituted or substituted C₂-C₅alkenylsuch as, for example, vinyl, allyl, 1-methyl propen-1-yl, butenyl, orpentenyl) or unsubstituted or substituted alkynyl (including but notlimited to unsubstituted or substituted C₂-C₅alkynyl such as acetylenyl,propargyl, butynyl, or pentynyl). Alternatively, R³ is unsubstituted orsubstituted aryl (including but not limited to monocyclic or bicyclicaryl) or unsubstituted or substituted arylalkyl (including but notlimited to monocyclic or bicyclic aryl linked to alkyl wherein alkylincludes but is not limited to CH₃, —CH₂CH₃, n-propyl, isopropyln-butyl, sec-butyl, and pentyl). In some other embodiments, R³ isunsubstituted or substituted heteroaryl, including but not limited tomonocyclic and bicyclic heteroaryl. Monocyclic heteroaryl R³ includesbut is not limited to pyrrolyl, thienyl, furyl, pyridinyl, pyranyl,pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, thiazolyl, pyrazolyl,and oxazolyl. Bicyclic heteroaryl R³ includes but is not limited tobenzothiophenyl, benzofuryl, indolyl, quinolinyl, isoquinolinyl,benzimidazolyl benzoxazolyl, benzothiazolyl, quinazolinyl, azaindolyl,pyrazolopyrimidinyl, purinyl, pyrrolo [1, 2-b]pyridazinyl,pyrrolopyrimidinyl, indazolyl, pyrazolylpyridinyl, imidazo[1,2-a]pyridinyl, and pyrrolo[1, 2-][1, 2, 4]triazinyl. The presentinvention also provides compounds of Formula I wherein R³ isunsubstituted or substituted heteroarylalkyl, including but not limitedto monocyclic and bicyclic heteroaryl as described above, that arelinked to alkyl, which in turn includes but is not limited to CH₃,—CH₂CH₃, n-propyl, isopropyl, n-butyl, sec-butyl, and pentyl. In someembodiments, R³ is unsubstituted or substituted cycloalkyl (includingbut not limited to cyclopropyl, cyclobutyl, and cyclopentyl) orunsubstituted or substituted heteroalkyl (non-limiting examples includeethoxymethyl, methoxymethyl, and diethylaminomethyl). In some furtherembodiments, R³ is unsubstituted or substituted heterocycloalkyl whichincludes but is not limited to pyrrolidinyl, tetrahydrofuranyl,piperidinyl, tetrahydropyranyl, thiazolidinyl, imidazolidinyl,morpholinyl, and piperazinyl. In yet other embodiments of the compoundsof Formula I, R³ is unsubstituted or substituted alkoxy including butnot limited to C₁-C₄alkoxy such as methoxy, ethoxy, propoxy or butoxy.R³ can also be unsubstituted or substituted heterocycloalkyloxy,including but not limited to 4-NH piperidin-1-yl-oxy, 4-methylpiperidin-1-yl-oxy, 4-ethyl piperidin-1-yl-oxy,4-isopropyl-piperidin-1-yl-oxy, and pyrrolidin-3-yl-oxy. In otherembodiments, R³ is unsubstituted or substituted amino, wherein thesubstituted amino includes but is not limited to dimethylamino,diethylamino, di-isopropyl amino, N-methyl N-ethyl amino, anddibutylamino. In some embodiments, R³ is unsubstituted or substitutedacyl, unsubstituted or substituted acyloxy, unsubstituted or substitutedC₁-C₄acyloxy, unsubstituted or substituted alkoxycarbonyl, unsubstitutedor substituted amido, or unsubstituted or substituted sulfonamido. Inother embodiments, R³ is halo, which is —I, —F, —Cl, or —Br. In someembodiments, R³ is selected from the group consisting of cyano, hydroxy,nitro, phosphate, urea, and carbonate. Also contemplated are R³ being—CH₃, —CH₂CH₃, n-propyl, isopropyl, n-butyl, tert-butyl, sec-butyl,pentyl, hexyl, heptyl, —OCH₃, —OCH₂CH₃, or —CF₃. In some embodiments R³can also be NR′R″ wherein R′ and R″ are taken together with the nitrogento form a cyclic moiety having from 3 to 8 ring atoms. The cyclic moietyso formed may further include one or more heteroatoms which are selectedfrom the group consisting of S, O, and N. The cyclic moiety so formed isunsubstituted or substituted, including but not limited to morpholinyl,azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, isothiazolidinyl1,2, dioxide, and thiomorpholinyl. Further non-limiting exemplary cyclicmoieites are the following:

The invention further provides a compound of Formula III:

or its pharmaceutically acceptable salts thereof wherein

-   -   W¹ is CR³, W² is C-benzoxazolyl substituted with R² and W³ is S;    -   W¹ is CR³, W² is C-benzoxazolyl substituted with R² and W³ is        CR⁵;    -   W¹ is N or NR³, W² is CR⁴, and W³ is C-benzoxazolyl substituted        with R²;    -   W¹ is CR³, W² is CR⁴, and W³ is C-benzoxazolyl substituted with        R²; or    -   W¹ is N or NR³, W² is NR⁴, and W³ is C-benzoxazolyl substituted        with R²;    -   X is N;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, alkyl,        heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,        aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,        heterocycloalkyloxy, amido, amino, acyl, acyloxy,        alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,        phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are taken        together with nitrogen to form a cyclic moiety.

In some embodiments of the compound of Formula III, the compound is:

and wherein W¹ is CR³ or NR³ and W² is CR⁴.

In another aspect, the invention provides a compound of Formula IV:

or its pharmaceutically acceptable salts thereof, wherein:

-   -   W¹ is N, NR³, CR³, or C═O; W² is N, NR⁴, CR⁴, or C═O; W³ is N,        NR⁵ or CR⁵; W⁴ is N, C═O or CR⁶, wherein no more than two N        atoms and no more than two C═O groups are adjacent;    -   W⁵ is N or CR⁷;    -   W⁶ is N or CR⁸;    -   W^(a) and W^(b) are independently N or CR⁹;    -   one of W^(c) and W^(d) is N, and the other is O, NR¹⁰, or S;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   or R³ and R⁴ taken together form a cyclic moiety;    -   R⁵, R⁶, R⁷ and R⁸ are independently hydrogen, alkyl,        heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,        aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,        heterocycloalkyloxy, amido, amino, acyl, acyloxy,        alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,        phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are taken        together with nitrogen to form a cyclic moiety;    -   R⁹ is alkyl or halo; and    -   R¹⁰ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

In some embodiments of the compound of Formula IV, W¹ is CR³, W² is CR⁴,W³ is CR³, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸; W¹ is N, W² is CR⁴, W³ isCR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸; or W¹ is CR³, W² is N, W³ isCR⁵, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸. In some embodiments of thecompound of Formula IV, W^(b) is N. In other embodiments, W^(a) is CR⁹and R⁹ is alkyl.

The invention also provides a compound of Formula V:

or its pharmaceutically acceptable salts thereof, wherein

-   -   W¹ is S, N, NR³ or CR³, W² is N or CR⁴, W³ is S, N or CR⁵, W⁴ is        N or C, and W⁷ is N or C, wherein no more than two N atoms and        no more than two C═O groups are adjacent;    -   W⁵ is N or CR⁷;    -   W⁶ is N or CR⁸;    -   W^(a) and W^(b) are independently N or CR⁹;    -   one of W^(c) and W^(d) is N, and the other is O, NR¹⁰, or S;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety,    -   or R³ and R⁴ taken together form a cyclic moiety;    -   R⁵, R⁷ and R⁸ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R⁹ is alkyl or halo; and    -   R¹⁰ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

In some embodiments of the compound of Formula V, W¹ is CR³, W² is CR⁴,W³ is N, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸. In other embodiments, W¹ isCR³, W² is CR⁴, W³ is N, W⁴ is N, W⁵ is CR⁷, and W⁶ is CR⁸. In otherembodiments, W¹ is CR³, W² is CR⁴, W³ is N, W⁴ is N, W⁵ is N, and W⁶ isCR⁸. In still other embodiments, W¹ is NR³, W² is CR⁴, W³ is N, W⁴ is C,W⁵ is CR⁷, and W⁶ is CR⁸. In other embodiments, W¹ is S, W² is CR⁴, W³is N, W⁴ is C, W⁵ is CR⁷, and W⁶ is CR⁸. In other embodiments, W¹ isCR³, W² is CR⁴, W³ is S, W⁴ is C, W⁵ is N, and W⁶ is N.

In some embodiments of the compound of Formula V, W^(b) is N. In otherembodiments, W^(a) is CR⁹ and R⁹ is alkyl.

The invention further provides compounds of Formula V-A and V-B:

or its pharmaceutically acceptable salts thereof, wherein

-   -   W¹ is CR³;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   and R³ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

Also provided herein are compounds of Formula V-C and V-D:

or its pharmaceutically acceptable salts thereof, wherein

-   -   W¹ is CR³;    -   W⁵ is N or CR⁷;    -   W^(a) and W^(b) are independently N or CR⁹;    -   one of W^(c) and W^(d) is N, and the other is O, NR¹⁰, or S;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R⁷ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R⁹ is alkyl or halo; and    -   R¹⁰ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

In some embodiments of the compound of Formula V-C or V-D, W^(b) is N.In other embodiments, W^(a) is CR⁹ and R⁹ is alkyl.

Also provided herein is a compound of Formula VI:

or its pharmaceutically acceptable salts thereof wherein

-   -   W¹ is CR³; W₂ is CR⁴;    -   W^(a) is CH or N;    -   R¹ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ is alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,        heterocycloalkyl, aryl, arylalkyl, heteroarylalkyl, alkoxy,        heterocycloalkyloxy, amido, amino, acyl, acyloxy,        alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,        phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are taken        together with nitrogen to form a cyclic moiety;    -   R⁴ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   or R³ and R⁴ taken together form a cyclic moiety; and    -   R¹⁰ and R¹¹ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

The invention further provides a compound of Formula VII:

-   -   or a pharmaceutically acceptable salt thereof, wherein    -   X₁ is CR³, NR³, or S;    -   X₂ is CR⁴, NR⁴, CR⁴        CR⁵, or CR⁴        NR⁵;    -   X₃ and X₄ are independently C or N;    -   X₅ is CR⁶, NR⁶, or S;    -   X₄ is CR⁷, NR⁷, CR⁷        CR⁸, or CR⁷        R⁸;    -   W^(a) and W^(b) are independently N or CR⁹;    -   one of W^(c) and W^(d) is N, and the other is O, NR¹⁰, or S;    -   R¹ and R² are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   R³ and R⁴ are independently hydrogen, alkyl, heteroalkyl,        alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido,        amino, acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety;    -   or R³ and R⁴ taken together form a cyclic moiety;    -   R⁵, R⁶, R⁷, and R⁸ are independently hydrogen, alkyl,        heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,        aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,        heterocycloalkyloxy, amido, amino, acyl, acyloxy,        alkoxycarbonyl, sulfonamido, halo, cyano, hydroxy, nitro,        phosphate, urea, carbonate, or NR′R″ wherein R′ and R″ are taken        together with nitrogen to form a cyclic moiety;    -   R⁹ is alkyl or halo; and    -   R¹⁰ is hydrogen, alkyl, heteroalkyl, alkenyl, alkynyl,        cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,        heteroarylalkyl, alkoxy, heterocycloalkyloxy, amido, amino,        acyl, acyloxy, alkoxycarbonyl, sulfonamido, halo, cyano,        hydroxy, nitro, phosphate, urea, carbonate, or NR′R″ wherein R′        and R″ are taken together with nitrogen to form a cyclic moiety.

In some embodiments of the compound of Formula VII, W^(b) is N. In otherembodiments, W^(a) is CR⁹ and R⁹ is alkyl.

In some embodiments, the compounds of the present invention exhibits oneor more functional characteristics disclosed herein. For example, one ormore subject compounds bind specifically to a PI3 kinase. In someembodiments, the IC50 of a subject compound for p110α, p110β, p110γ, orp110δ is less than about 1 μM, less than about 100 nM, less than about50 nM, less than about 10 nM, less than about 1 nM, less than about 0.5nM, less than about 100 pM, or less than about 50 pM.

In some embodiments, one or more of the subject compound may selectivelyinhibit one or more members of type I or class I phosphatidylinositol3-kinases (PI3-kinase) with an IC50 value of about 100 nM, 50 nM, 10 nM,5 nM, 100 pM, 10 pM or 1 pM, or less as measured in an in vitro kinaseassay.

In some embodiments, one or more of the subject compound may selectivelyinhibit one or two members of type I or class I phosphatidylinositol3-kinases (PI3-kinase) consisting of PI3-kinase α, PI3-kinase β,PI3-kinase γ, and PI3-kinase δ. In some aspects, some of the subjectcompounds selectively inhibit PI3-kinase α as compared to all other typeI PI3-kinases. In other aspects, some of the subject compoundsselectively inhibit PI3-kinase α and PI3-kinase γ as compared to therest of the type I PI3-kinases. In yet other aspects, some of thesubject compounds selectively inhibit PI3-kinase α and PI3-kinase β ascompared to the rest of the type I PI3-kinases. In still yet some otheraspects, some of the subject compounds selectively inhibit PI3-kinase αand PI3-kinase δ as compared to the rest of the type I PI3-kinases. Instill yet some other aspects, some of the subject compounds selectivelyinhibit PI3-kinase δ and PI3-kinase β as compared to the rest of thetype I PI3-kinases, or selectively inhibit PI3-kinase δ and PI3-kinase αas compared to the rest of the type I PI3-kinases, or selectivelyinhibit PI3-kinase α and PI3-kinase γ as compared to the rest of thetype I PI3-kinases, or selectively inhibit PI3-kinase γ and PI3-kinase βas compared to the rest of the type I PI3-kinases.

In yet another aspect, an inhibitor that selectively inhibits one ormore members of type I PI3-kinases, or an inhibitor that selectivelyinhibits one or more type I PI3-kinase mediated signaling pathways,alternatively can be understood to refer to a compound that exhibits a50% inhibitory concentration (IC50) with respect to a given type IPI3-kinase, that is at least at least 10-fold, at least 20-fold, atleast 50-fold, at least 100-fold, at least 1000-fold, at least10,100-fold, or lower, than the inhibitor's IC50 with respect to therest of the other type I PI3-kinases.

The chemical entities described herein can be synthesized according toone or more illustrative schemes herein and/or techniques known in theart.

Unless specified to the contrary, the reactions described herein takeplace at atmospheric pressure, generally within a temperature range from−10° C. to 200° C. Further, except as otherwise specified, reactiontimes and conditions are intended to be approximate, e.g., taking placeat about atmospheric pressure within a temperature range of about −10°C. to about 110° C. over a period of about 1 to about 24 hours:reactions left to run overnight average a period of about 16 hours.

In general, compounds of the invention may be prepared by the followingreaction schemes:

The compounds of the invention may be synthesized via a reaction schemerepresented generally in Schemes A and B. The synthesis proceeds viacoupling a compound of Formula A with a compound of Formula B to yield acompound of Formula C. The coupling step is typically catalyzed by usinga palladium catalyst, including but not limited to palladium tetrakis(triphenylphosphine). The coupling is generally performed in thepresence of a suitable base, a nonlimiting example being sodiumcarbonate. One example of a suitable solvent for the reaction is aqueousdioxane.

A compound of Formula A for use in Scheme A has a structure of FormulaA, wherein T₁ is halo including bromo, chloro, fluoro, and iodo, andwherein the remaining substituents are defined for Formulas I and II ofcompounds of the invention. For boronic acids and acid derivatives asdepicted in Formula B, X is either O or S, and the benzoxazole orbenzothiazole moiety can be attached at the 4-, 5-, 6- or 7-position.

For a compound of Formula B, G is hydrogen or R_(G1), wherein R_(G1) isalkyl, alkenyl, or aryl. Alternatively, B(OG)₂ is taken together to forma 5- or 6-membered cyclic moiety. In some embodiments, the compound ofFormula B is a compound having a structure of Formula E:

wherein G is H or R_(G1); R_(G1) is alkyl, alkenyl, or aryl.Alternatively, B(OG)₂ is taken together to form a 5- or 6-memberedcyclic moiety and R_(G2) is H, tert-butyl carbamate, or acyl.

Scheme C depicts an exemplary scheme for synthesizing a compound ofFormula B′ or, optionally, Formula B″ for use in Reaction Scheme C. M isa benzoxazolyl or benzothiazolyl moiety as described by Formula B. Thisreaction proceeds via reacting a compound of Formula D with a trialkylborate or a boronic acid derivative to produce a compound of Formula B′.The trialkyl borate includes but is not limited to triisopropyl borateand the boronic acid derivative includes but is not limited tobis(pinacolato)diboron. The reaction typically is run in the presence ofa base, a nonlimiting example being potassium acetate. The reaction maybe run in a solvent such as dioxane or tetrahydrofuran.

A compound of Formula D for use in Scheme C is a compound wherein T₂ ishalo or another leaving group, and M is as defined above. The compoundof Formula B′ may further be converted to a compound of Formula B″ bytreatment with an acid such as hydrochloric acid.

Some exemplary compounds of Formula B that can be synthesized via SchemeC include but are not limited to compounds of the following formulae:

Where desired, deprotection of a substituent (e.g., removal of Bocprotection from an amino substituent) on the benzoxazolyl moiety (i.e.M₁ of Formula C) is performed after coupling the compound of Formula Bto the compound of Formula A.

Some exemplary compounds with such protecting groups, include but arenot limited to compounds of the following formulae:

The following Reaction Schemes illustrate the preparation of severalcompounds of the invention.

The invention provides pharmaceutical compositions comprising one ormore compounds of the present invention.

In some embodiments, the invention provides pharmaceutical compositionsfor the treatment of disorders such as hyperproliferative disorderincluding but not limited to cancer such as acute myeloid leukemia,thymus, brain, lung, squamous cell, skin, eye, retinoblastoma,intraocular melanoma, oral cavity and oropharyngeal, bladder, gastric,stomach, pancreatic, bladder, breast, cervical, head, neck, renal,kidney, liver, ovarian, prostate, colorectal, esophageal, testicular,gynecological, thyroid, CNS, PNS, AIDS-Related (e.g., Lymphoma andKaposi's Sarcoma) or Viral-Induced cancer. In some embodiments, saidpharmaceutical composition is for the treatment of a non-canceroushyperproliferative disorder such as benign hyperplasia of the skin (e.g., psoriasis), restenosis, or prostate (e. g., benign prostatichypertrophy (BPH)).

In some embodiments, the invention provides pharmaceutical compositionsfor treating diseases or conditions related to an undesirable,over-active, harmful or deleterious immune response in a mammal. Suchundesirable immune response can be associated with or result in, e.g.,asthma, emphysema, bronchitis, psoriasis, allergy, anaphylaxsis,auto-immune diseases, rheumatoid arthritis, graft versus host disease,transplantation rejection, lung injuries, and lupus erythematosus. Thepharmaceutical compositions of the present invention can be used totreat other respiratory diseases including but not limited to diseasesaffecting the lobes of lung, pleural cavity, bronchial tubes, trachea,upper respiratory tract, or the nerves and muscle for breathing. Thecompositions of the invention can be further used to treat multiorganfailure.

The invention also provides compositions for the treatment of liverdiseases (including diabetes), pancreatitis or kidney disease (includingproliferative glomerulonephritis and diabetes-induced renal disease) orpain in a mammal.

The invention also provides compositions for the treatment of spermmotility. The invention further provides compositions for the treatmentof neurological or neurodegenerative diseases including, but not limitedto, Alzheimer's disease, Huntington's disease, CNS trauma, and stroke.

The invention further provides a composition for the prevention ofblastocyte implantation in a mammal.

The invention also relates to a composition for treating a diseaserelated to vasculogenesis or angiogenesis in a mammal which can manifestas tumor angiogenesis, chronic inflammatory disease such as rheumatoidarthritis, inflammatory bowel disease, atherosclerosis, skin diseasessuch as psoriasis, eczema, and scleroderma, diabetes, diabeticretinopathy, retinopathy of prematurity, age-related maculardegeneration, hemangioma, glioma, melanoma, Kaposi's sarcoma andovarian, breast, lung, pancreatic, prostate, colon and epidermoidcancer.

The invention further provides compositions for the treatment ofdisorders involving platelet aggregation or platelet adhesion, includingbut not limited to Bernard-Soulier syndrome, Glanzmann's thrombasthenia,Scott's syndrome, von Willebrand disease, Hermansky-Pudlak Syndrome, andGray platelet syndrome.

In some embodiments, compositions are provided for treating a diseasewhich is skeletal muscle atrophy, skeletal muscle hypertrophy, leukocyterecruitment in cancer tissue, invasion metastasis, melanoma, Kaposi'ssarcoma, acute and chronic bacterial and viral infections, sepsis,glomerulo sclerosis, glomerulo, nephritis, or progressive renalfibrosis.

The subject pharmaceutical compositions are typically formulated toprovide a therapeutically effective amount of a compound of the presentinvention as the active ingredient, or a pharmaceutically acceptablesalt, ester, prodrug, solvate, hydrate or derivative thereof. Wheredesired, the pharmaceutical compositions contain pharmaceuticallyacceptable salt and/or coordination complex thereof, and one or morepharmaceutically acceptable excipients, carriers, including inert soliddiluents and fillers, diluents, including sterile aqueous solution andvarious organic solvents, permeation enhancers, solubilizers andadjuvants.

The subject pharmaceutical compositions can be administered alone or incombination with one or more other agents, which are also typicallyadministered in the form of pharmaceutical compositions. Where desired,the subject compounds and other agent(s) may be mixed into a preparationor both components may be formulated into separate preparations to usethem in combination separately or at the same time.

In some embodiments, the concentration of one or more of the compoundsprovided in the pharmaceutical compositions of the present invention isless than 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%,16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%,0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%,0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%,0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%,0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w, w/v or v/v.

In some embodiments, the concentration of one or more of the compoundsof the present invention is greater than 90%, 80%, 70%, 60%, 50%, 40%,30%, 20%, 19.75%, 19.50%, 19.25% 19%, 18.75%, 18.50%, 18.25% 18%,17.75%, 17.50%, 17.25% 17%, 16.75%, 16.50%, 16.25% 16%, 15.75%, 15.50%,15.25% 15%, 14.75%, 14.50%, 14.25% 14%, 13.75%, 13.50%, 13.25% 13%,12.75%, 12.50%, 12.25% 12%, 11.75%, 11.50%, 11.25% 11%, 10.75%, 10.50%,10.25% 10%, 9.75%, 9.50%, 9.25% 9%, 8.75%, 8.50%, 8.25% 8%, 7.75%,7.50%, 7.25% 7%, 6.75%, 6.50%, 6.25% 6%, 5.75%, 5.50%, 5.25% 5%, 4.75%,4.50%, 4.25%, 4%, 3.75%, 3.50%, 3.25%, 3%, 2.75%, 2.50%, 2.25%, 2%,1.75%, 1.50%, 125%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%,0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%,0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%,0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w,w/v, or v/v.

In some embodiments, the concentration of one or more of the compoundsof the present invention is in the range from approximately 0.0001% toapproximately 50%, approximately 0.001% to approximately 40%,approximately 0.01% to approximately 30%, approximately 0.02% toapproximately 29%, approximately 0.03% to approximately 28%,approximately 0.04% to approximately 27%, approximately 0.05% toapproximately 26%, approximately 0.06% to approximately 25%,approximately 0.07% to approximately 24%, approximately 0.08% toapproximately 23%, approximately 0.09% to approximately 22%,approximately 0.1% to approximately 21%, approximately 0.2% toapproximately 20%, approximately 0.3% to approximately 19%,approximately 0.4% to approximately 18%, approximately 0.5% toapproximately 17%, approximately 0.6% to approximately 16%,approximately 0.7% to approximately 15%, approximately 0.8% toapproximately 14%, approximately 0.9% to approximately 12%,approximately 1% to approximately 10% w/w, w/v or v/v, v/v.

In some embodiments, the concentration of one or more of the compoundsof the present invention is in the range from approximately 0.001% toapproximately 10%, approximately 0.01% to approximately 5%,approximately 0.02% to approximately 4.5%, approximately 0.03% toapproximately 4%, approximately 0.04% to approximately 3.5%,approximately 0.05% to approximately 3%, approximately 0.06% toapproximately 2.5%, approximately 0.07% to approximately 2%,approximately 0.08% to approximately 1.5%, approximately 0.09% toapproximately 1%, approximately 0.1% to approximately 0.9% w/w, w/v orv/v.

In some embodiments, the amount of one or more of the compounds of thepresent invention is equal to or less than 10 g, 9.5 g, 9.0 g, 8.5 g,8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g,3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g,0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25g, 0.2 g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g,0.03 g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g,0.004 g, 0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006g, 0.0005 g, 0.0004 g, 0.0003 g, 0.0002 g, or 0.0001 g.

In some embodiments, the amount of one or more of the compounds of thepresent invention is more than 0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g,0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.001 g, 0.0015 g,0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004 g, 0.0045 g, 0.005 g, 0.0055g, 0.006 g, 0.0065 g, 0.007 g, 0.0075 g, 0.008 g, 0.0085 g, 0.009 g,0.0095 g, 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g, 0.04 g,0.045 g, 0.05 g, 0.055 g, 0.06 g, 0.065 g, 0.07 g, 0.075 g, 0.08 g,0.085 g, 0.09 g, 0.095 g, 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35 g,0.4 g, 0.45 g, 0.5 g, 0.55 g, 0.6 g, 0.65 g, 0.7 g, 0.75 g, 0.8 g, 0.85g, 0.9 g, 0.95 g, 1 g, 1.5 g, 2 g, 2.5, 3 g, 3.5, 4 g, 4.5 g, 5 g, 5.5g, 6 g, 6.5 g, 7 g, 7.5 g, 8 g, 8.5 g, 9 g, 9.5 g, or 10 g.

In some embodiments, the amount of one or more of the compounds of thepresent invention is in the range of 0.0001-10 g, 0.0005-9 g, 0.001-8 g,0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g.

The compounds according to the invention are effective over a widedosage range. For example, in the treatment of adult humans, dosagesfrom 0.01 to 1000 mg, from 0.5 to 100 mg, from 1 to 50 mg per day, andfrom 5 to 40 mg per day are examples of dosages that may be used. Anexemplary dosage is 10 to 30 mg per day. The exact dosage will dependupon the route of administration, the form in which the compound isadministered, the subject to be treated, the body weight of the subjectto be treated, and the preference and experience of the attendingphysician.

A pharmaceutical composition of the present invention typically containsan active ingredient (e.g., a compound of the present invention or apharmaceutically acceptable salt and/or coordination complex thereof,and one or more pharmaceutically acceptable excipients, carriers,including but not limited inert solid diluents and fillers, diluents,sterile aqueous solution and various organic solvents, permeationenhancers, solubilizers and adjuvants.

Described below are non-limiting exemplary pharmaceutical compositionsand methods for preparing the same.

Pharmaceutical Compositions for Oral Administration

In some embodiments, the invention provides a pharmaceutical compositionfor oral administration containing a compound of the present invention,and a pharmaceutical excipient suitable for oral administration.

In some embodiments, the invention provides a solid pharmaceuticalcomposition for oral administration containing: (i) an effective amountof a compound of the present invention; optionally (ii) an effectiveamount of a second agent; and (iii) a pharmaceutical excipient suitablefor oral administration. In some embodiments, the composition furthercontains: (iv) an effective amount of a third agent.

In some embodiments, the pharmaceutical composition may be a liquidpharmaceutical composition suitable for oral consumption. Pharmaceuticalcompositions of the invention suitable for oral administration can bepresented as discrete dosage forms, such as capsules, cachets, ortablets, or liquids or aerosol sprays each containing a predeterminedamount of an active ingredient as a powder or in granules, a solution,or a suspension in an aqueous or non-aqueous liquid, an oil-in-wateremulsion, or a water-in-oil liquid emulsion. Such dosage forms can beprepared by any of the methods of pharmacy, but all methods include thestep of bringing the active ingredient into association with thecarrier, which constitutes one or more necessary ingredients. Ingeneral, the compositions are prepared by uniformly and intimatelyadmixing the active ingredient with liquid carriers or finely dividedsolid carriers or both, and then, if necessary, shaping the product intothe desired presentation. For example, a tablet can be prepared bycompression or molding, optionally with one or more accessoryingredients. Compressed tablets can be prepared by compressing in asuitable machine the active ingredient in a free-flowing form such aspowder or granules, optionally mixed with an excipient such as, but notlimited to, a binder, a lubricant, an inert diluent, and/or a surfaceactive or dispersing agent. Molded tablets can be made by molding in asuitable machine a mixture of the powdered compound moistened with aninert liquid diluent.

This invention further encompasses anhydrous pharmaceutical compositionsand dosage forms comprising an active ingredient, since water canfacilitate the degradation of some compounds. For example, water may beadded (e.g., 5%) in the pharmaceutical arts as a means of simulatinglong-term storage in order to determine characteristics such asshelf-life or the stability of formulations over time. Anhydrouspharmaceutical compositions and dosage forms of the invention can beprepared using anhydrous or low moisture containing ingredients and lowmoisture or low humidity conditions. Pharmaceutical compositions anddosage forms of the invention which contain lactose can be madeanhydrous if substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected. An anhydrouspharmaceutical composition may be prepared and stored such that itsanhydrous nature is maintained. Accordingly, anhydrous compositions maybe packaged using materials known to prevent exposure to water such thatthey can be included in suitable formulary kits. Examples of suitablepackaging include, but are not limited to, hermetically sealed foils,plastic or the like, unit dose containers, blister packs, and strippacks.

An active ingredient can be combined in an intimate admixture with apharmaceutical carrier according to conventional pharmaceuticalcompounding techniques. The carrier can take a wide variety of formsdepending on the form of preparation desired for administration. Inpreparing the compositions for an oral dosage form, any of the usualpharmaceutical media can be employed as carriers, such as, for example,water, glycols, oils, alcohols, flavoring agents, preservatives,coloring agents, and the like in the case of oral liquid preparations(such as suspensions, solutions, and elixirs) or aerosols; or carrierssuch as starches, sugars, micro-crystalline cellulose, diluents,granulating agents, lubricants, binders, and disintegrating agents canbe used in the case of oral solid preparations, in some embodimentswithout employing the use of lactose. For example, suitable carriersinclude powders, capsules, and tablets, with the solid oralpreparations. If desired, tablets can be coated by standard aqueous ornonaqueous techniques.

Binders suitable for use in pharmaceutical compositions and dosage formsinclude, but are not limited to, corn starch, potato starch, or otherstarches, gelatin, natural and synthetic gums such as acacia, sodiumalginate, alginic acid, other alginates, powdered tragacanth, guar gum,cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate,carboxymethyl cellulose calcium, sodium carboxymethyl cellulose),polyvinyl pyrrolidone, methyl cellulose, pre-gelatinized starch,hydroxypropyl methyl cellulose, microcrystalline cellulose, and mixturesthereof.

Examples of suitable fillers for use in the pharmaceutical compositionsand dosage forms disclosed herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.

Disintegrants may be used in the compositions of the invention toprovide tablets that disintegrate when exposed to an aqueousenvironment. Too much of a disintegrant may produce tablets which maydisintegrate in the bottle. Too little may be insufficient fordisintegration to occur and may thus alter the rate and extent ofrelease of the active ingredient(s) from the dosage form. Thus, asufficient amount of disintegrant that is neither too little nor toomuch to detrimentally alter the release of the active ingredient(s) maybe used to form the dosage forms of the compounds disclosed herein. Theamount of disintegrant used may vary based upon the type of formulationand mode of administration, and may be readily discernible to those ofordinary skill in the art. About 0.5 to about 15 weight percent ofdisintegrant, or about 1 to about 5 weight percent of disintegrant, maybe used in the pharmaceutical composition. Disintegrants that can beused to form pharmaceutical compositions and dosage forms of theinvention include, but are not limited to, agar-agar, alginic acid,calcium carbonate, microcrystalline cellulose, croscarmellose sodium,crospovidone, polacrilin potassium, sodium starch glycolate, potato ortapioca starch, other starches, pre-gelatinized starch, other starches,clays, other algins, other celluloses, gums or mixtures thereof.

Lubricants which can be used to form pharmaceutical compositions anddosage forms of the invention include, but are not limited to, calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethylaureate, agar, ormixtures thereof. Additional lubricants include, for example, a syloidsilica gel, a coagulated aerosol of synthetic silica, or mixturesthereof. A lubricant can optionally be added, in an amount of less thanabout 1 weight percent of the pharmaceutical composition.

When aqueous suspensions and/or elixirs are desired for oraladministration, the essential active ingredient therein may be combinedwith various sweetening or flavoring agents, coloring matter or dyesand, if so desired, emulsifying and/or suspending agents, together withsuch diluents as water, ethanol, propylene glycol, glycerin and variouscombinations thereof.

The tablets can be uncoated or coated by known techniques to delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearate canbe employed. Formulations for oral use can also be presented as hardgelatin capsules wherein the active ingredient is mixed with an inertsolid diluent, for example, calcium carbonate, calcium phosphate orkaolin, or as soft gelatin capsules wherein the active ingredient ismixed with water or an oil medium, for example, peanut oil, liquidparaffin or olive oil.

Surfactant which can be used to form pharmaceutical compositions anddosage forms of the invention include, but are not limited to,hydrophilic surfactants, lipophilic surfactants, and mixtures thereof.That is, a mixture of hydrophilic surfactants may be employed, a mixtureof lipophilic surfactants may be employed, or a mixture of at least onehydrophilic surfactant and at least one lipophilic surfactant may beemployed.

A suitable hydrophilic surfactant may generally have an HLB value of atleast 10, while suitable lipophilic surfactants may generally have anHLB value of or less than about 10. An empirical parameter used tocharacterize the relative hydrophilicity and hydrophobicity of non-ionicamphiphilic compounds is the hydrophilic-lipophilic balance (“HLB”value). Surfactants with lower HLB values are more lipophilic orhydrophobic, and have greater solubility in oils, while surfactants withhigher HLB values are more hydrophilic, and have greater solubility inaqueous solutions. Hydrophilic surfactants are generally considered tobe those compounds having an HLB value greater than about 10, as well asanionic, cationic, or zwitterionic compounds for which the HLB scale isnot generally applicable. Similarly, lipophilic (i.e., hydrophobic)surfactants are compounds having an HLB value equal to or less thanabout 10. However, HLB value of a surfactant is merely a rough guidegenerally used to enable formulation of industrial, pharmaceutical andcosmetic emulsions.

Hydrophilic surfactants may be either ionic or non-ionic. Suitable ionicsurfactants include, but are not limited to, alkylammonium salts;fusidic acid salts; fatty acid derivatives of amino acids,oligopeptides, and polypeptides; glyceride derivatives of amino acids,oligopeptides, and polypeptides; lecithins and hydrogenated lecithins;lysolecithins and hydrogenated lysolecithins; phospholipids andderivatives thereof; lysophospholipids and derivatives thereof;carnitine fatty acid ester salts; salts of alkylsulfates; fatty acidsalts; sodium docusate; acylactylates; mono- and di-acetylated tartaricacid esters of mono- and di-glycerides; succinylated mono- anddi-glycerides; citric acid esters of mono- and di-glycerides; andmixtures thereof.

Within the aforementioned group, ionic surfactants include, by way ofexample: lecithins, lysolecithin, phospholipids, lysophospholipids andderivatives thereof; carnitine fatty acid ester salts; salts ofalkylsulfates; fatty acid salts; sodium docusate; acylactylates; mono-and di-acetylated tartaric acid esters of mono- and di-glycerides;succinylated mono- and di-glycerides; citric acid esters of mono- anddi-glycerides; and mixtures thereof.

Ionic surfactants may be the ionized forms of lecithin, lysolecithin,phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol,phosphatidic acid, phosphatidylserine, lysophosphatidylcholine,lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidicacid, lysophosphatidylserine, PEG-phosphatidylethanolamine,PVP-phosphatidylethanolamine, lactylic esters of fatty acids,stearoyl-2-lactylate, stearoyl lactylate, succinylated monoglycerides,mono/diacetylated tartaric acid esters of mono/diglycerides, citric acidesters of mono/diglycerides, cholylsarcosine, caproate, caprylate,caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate,linolenate, stearate, lauryl sulfate, teracecyl sulfate, docusate,lauroyl carnitines, palmitoyl carnitines, myristoyl carnitines, andsalts and mixtures thereof.

Hydrophilic non-ionic surfactants may include, but not limited to,alkylglucosides; alkylmaltosides; alkylthioglucosides; laurylmacrogolglycerides; polyoxyalkylene alkyl ethers such as polyethyleneglycol alkyl ethers; polyoxyalkylene alkylphenols such as polyethyleneglycol alkyl phenols; polyoxyalkylene alkyl phenol fatty acid esterssuch as polyethylene glycol fatty acids monoesters and polyethyleneglycol fatty acids diesters; polyethylene glycol glycerol fatty acidesters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fattyacid esters such as polyethylene glycol sorbitan fatty acid esters;hydrophilic transesterification products of a polyol with at least onemember of the group consisting of glycerides, vegetable oils,hydrogenated vegetable oils, fatty acids, and sterols; polyoxyethylenesterols, derivatives, and analogues thereof; polyoxyethylated vitaminsand derivatives thereof; polyoxyethylene-polyoxypropylene blockcopolymers; and mixtures thereof; polyethylene glycol sorbitan rattyacid esters and hydrophilic transesterification products or a polyolwith at least one member of the group consisting of triglycerides,vegetable oils, and hydrogenated vegetable oils. The polyol may beglycerol, ethylene glycol, polyethylene glycol, sorbitol, propyleneglycol, pentaerythritol, or a saccharide.

Other hydrophilic-non-ionic surfactants include, without limitation,PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate,PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate,PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryllaurate, PEG-40 palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenatedcastor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6caprate/caprylate glycerides, PEG-8 caprate/caprylate glycerides,polyglyceryl-10 laurate, PEG-30 cholesterol, PEG-25 phyto sterol, PEG-30soya sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitanlaurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearylether, tocopheryl PEG-100 succinate, PEG-24 cholesterol,polyglyceryl-10oleate, Tween 40, Tween 60, sucrose monostearate, sucrosemonolaurate, sucrose monopalmitate, PEG 10-100 nonyl phenol series, PEG15-100 octyl phenol series, and poloxamers.

Suitable lipophilic surfactants include, by way of example only: fattyalcohols; glycerol fatty acid esters; acetylated glycerol fatty acidesters; lower alcohol fatty acids esters; propylene glycol fatty acidesters; sorbitan ratty acid esters; polyethylene glycol sorbitan rattyacid esters; sterols and sterol derivatives; polyoxyethylated sterolsand sterol derivatives; polyethylene glycol alkyl ethers; sugar esters;sugar ethers; lactic acid derivatives of mono- and di-glycerides;hydrophobic transesterification products of a polyol with at least onemember of the group consisting of glycerides, vegetable oils,hydrogenated vegetable oils, fatty acids and sterols; oil-solublevitamins/vitamin derivatives; and mixtures thereof. Within this group,preferred lipophilic surfactants include glycerol fatty acid esters,propylene glycol fatty acid esters, and mixtures thereof, or arehydrophobic transesterification products of a polyol with at least onemember of the group consisting of vegetable oils, hydrogenated vegetableoils, and triglycerides.

In one embodiment, the composition may include a solubilizer to ensuregood solubilization and/or dissolution of the compound of the presentinvention and to minimize precipitation of the compound of the presentinvention. This can be especially important for compositions fornon-oral use, e.g., compositions for injection. A solubilizer may alsobe added to increase the solubility of the hydrophilic drug and/or othercomponents, such as surfactants, or to maintain the composition as astable or homogeneous solution or dispersion.

Examples of suitable solubilizers include, but are not limited to, thefollowing: alcohols and polyols, such as ethanol, isopropanol, butanol,benzyl alcohol, ethylene glycol, propylene glycol, butanediols andisomers thereof, glycerol, pentaerythritol, sorbitol, mannitol,transcutol, dimethyl isosorbide, polyethylene glycol, polypropyleneglycol, polyvinylalcohol, hydroxypropyl methylcellulose and othercellulose derivatives, cyclodextrins and cyclodextrin derivatives;ethers of polyethylene glycols having an average molecular weight ofabout 200 to about 6000, such as tetrahydrofurfuryl alcohol PEG ether(glycofurol) or methoxy PEG; amides and other nitrogen-containingcompounds such as 2-pyrrolidone, 2-piperidone, .epsilon.-caprolactam,N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone,N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrolidone; esterssuch as ethyl propionate, tributylcitrate, acetyl triethylcitrate,acetyl tributyl citrate, triethylcitrate, ethyl oleate, ethyl caprylate,ethyl butyrate, triacetin, propylene glycol nmonoacetate, propyleneglycol diacetate, e-caprolactone and isomers thereof, δ-valerolactoneand isomers thereof, β-butyrolactone and isomers thereof; and othersolubilizers known in the art, such as dimethyl acetamide, dimethylisosorbide, N-methyl pyrrolidones, monooctanoin, diethylene glycolmonoethyl ether, and water.

Mixtures of solubilizers may also be used. Examples include, but notlimited to, triacetin, triethylcitrate, ethyl oleate, ethyl caprylate,dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone,polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropylcyclodextrins, ethanol, polyethylene glycol 200-100, glycofurol,transcutol, propylene glycol, and dimethyl isosorbide. Particularlypreferred solubilizers include sorbitol, glycerol, triacetin, ethylalcohol, PEG-400, glycofurol and propylene glycol.

The amount of solubilizer that can be included is not particularlylimited. The amount of a given solubilizer may be limited to abioacceptable amount, which may be readily determined by one of skill inthe art. In some circumstances, it may be advantageous to includeamounts of solubilizers far in excess of bioacceptable amounts, forexample to maximize the concentration of the drug, with excesssolubilizer removed prior to providing the composition to a patientusing conventional techniques, such as distillation or evaporation.Thus, if present, the solubilizer can be in a weight ratio of 10%, 25%,50%, 100%, or up to about 200% by weight, based on the combined weightof the drug, and other excipients. If desired, very small amounts ofsolubilizer may also be used, such as 5%, 2%, 1% or even less.Typically, the solubilizer may be present in an amount of about 1% toabout 100%, more typically about 5% to about 25% by weight.

The composition can further include one or more pharmaceuticallyacceptable additives and excipients. Such additives and excipientsinclude, without limitation, detackifiers, anti-foaming agents,buffering agents, polymers, antioxidants, preservatives, chelatingagents, viscomodulators, tonicifiers, flavorants, colorants, odorants,opacitiers, suspending agents, binders, fillers, plasticizers,lubricants, and mixtures thereof.

In addition, an acid or a base may be incorporated into the compositionto facilitate processing, to enhance stability, or for other reasons.Examples of pharmaceutically acceptable bases include amino acids, aminoacid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide,sodium hydrogen carbonate, aluminum hydroxide, calcium carbonate,magnesium hydroxide, magnesium aluminum silicate, synthetic aluminumsilicate, synthetic hydrocalcite, magnesium aluminum hydroxide,diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine,triethylamine, triisopropanolamine, trimethylamine,tris(hydroxymethyl)aminomethane (TRIS) and the like. Also suitable arebases that are salts of a pharmaceutically acceptable acid, such asacetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonicacid, amino acids, ascorbic acid, benzoic acid, boric acid, butyricacid, carbonic acid, citric acid, fatty acids, formic acid, fumaricacid, gluconic acid, hydroquinosulfonic acid, isoascorbic acid, lacticacid, maleic acid, oxalic acid, para-bromophenylsulfonic acid, propionicacid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinicacid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonicacid, uric acid, and the like. Salts of polyprotic acids, such as sodiumphosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphatecan also be used. When the base is a salt, the cation can be anyconvenient and pharmaceutically acceptable cation, such as ammonium,alkali metals, alkaline earth metals, and the like. Example may include,but not limited to, sodium, potassium, lithium, magnesium, calcium andammonium.

Suitable acids are pharmaceutically acceptable organic or inorganicacids. Examples of suitable inorganic acids include hydrochloric acid,hydrobromic acid, hydriodic acid, sulfuric acid, nitric acid, boricacid, phosphoric acid, and the like. Examples of suitable organic acidsinclude acetic acid, acrylic acid, adipic acid, alginic acid,alkanesulfonic acids, amino acids, ascorbic acid, benzoic acid, boricacid, butyric acid, carbonic acid, citric acid, fatty acids, formicacid, fumaric acid, gluconic acid, hydroquinosulfonic acid, isoascorbicacid, lactic acid, malcic acid, methanesulfonic acid, oxalic acid,para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid,salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid,thioglycolic acid, toluenesulfonic acid, uric acid and the like.

Pharmaceutical Compositions for Injection.

In some embodiments, the invention provides a pharmaceutical compositionfor injection containing a compound of the present invention and apharmaceutical excipient suitable for injection. Components and amountsof agents in the compositions are as described herein.

The forms in which the novel compositions of the present invention maybe incorporated for administration by injection include aqueous or oilsuspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, orpeanut oil as well as elixirs, mannitol, dextrose, or a sterile aqueoussolution, and similar pharmaceutical vehicles.

Aqueous solutions in saline are also conventionally used for injection.Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, and thelike (and suitable mixtures thereof), cyclodextrin derivatives, andvegetable oils may also be employed. The proper fluidity can bemaintained, for example, by the use of a coating, such as lecithin, forthe maintenance of the required particle size in the case of dispersionand by the use of surfactants. The prevention of the action ofmicroorganisms can be brought about by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, thimerosal, and the like.

Sterile injectable solutions are prepared by incorporating the compoundof the present invention in the required amount in the appropriatesolvent with various other ingredients as enumerated above, as required,followed by filtered sterilization. Generally, dispersions are preparedby incorporating the various sterilized active ingredients into asterile vehicle which contains the basic dispersion medium and therequired other ingredients from those enumerated above. In the case ofsterile powders for the preparation of sterile injectable solutions,certain desirable methods of preparation are vacuum-drying andfreeze-drying techniques which yield a powder of the active ingredientplus any additional desired ingredient from a previouslysterile-filtered solution thereof.

Pharmaceutical Compositions for Topical (e.g., Transdermal) Delivery.

In some embodiments, the invention provides a pharmaceutical compositionfor transdermal delivery containing a compound of the present inventionand a pharmaceutical excipient suitable for transdermal delivery.

Compositions of the present invention can be formulated intopreparations in solid, semi-solid, or liquid forms suitable for local ortopical administration, such as gels, water soluble jellies, creams,lotions, suspensions, foams, powders, slurries, ointments, solutions,oils, pastes, suppositories, sprays, emulsions, saline solutions,dimethylsulfoxide (DMSO)-based solutions. In general, carriers withhigher densities are capable of providing an area with a prolongedexposure to the active ingredients. In contrast, a solution formulationmay provide more immediate exposure of the active ingredient to thechosen area.

The pharmaceutical compositions also may comprise suitable solid or gelphase carriers or excipients, which are compounds that allow increasedpenetration of, or assist in the delivery of, therapeutic moleculesacross the stratum corneum permeability barrier of the skin. There aremany of these penetration-enhancing molecules known to those trained inthe art of topical formulation. Examples of such carriers and excipientsinclude, but are not limited to, humectants (e.g., urea), glycols (e.g.,propylene glycol), alcohols (e.g., ethanol), fatty acids (e.g., oleicacid), surfactants (e.g., isopropyl myristate and sodium laurylsulfate), pyrrolidones, glycerol monolaurate, sulfoxides, terpenes(e.g., menthol), amines, amides, alkanes, alkanols, water, calciumcarbonate, calcium phosphate, various sugars, starches, cellulosederivatives, gelatin, and polymers such as polyethylene glycols.

Another exemplary formulation for use in the methods of the presentinvention employs transdermal delivery devices (“patches”). Suchtransdermal patches may be used to provide continuous or discontinuousinfusion of a compound of the present invention in controlled amounts,either with or without another agent.

The construction and use of transdermal patches for the delivery ofpharmaceutical agents is well known in the art. See, e.g., U.S. Pat.Nos. 5,023,252, 4,992,445 and 5,001,139. Such patches may be constructedfor continuous, pulsatile, or on demand delivery of pharmaceuticalagents.

Pharmaceutical Compositions for Inhalation.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra. Preferably the compositions are administered by the oral or nasalrespiratory route for local or systemic effect. Compositions inpreferably pharmaceutically acceptable solvents may be nebulized by useof inert gases. Nebulized solutions may be inhaled directly from thenebulizing device or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine.Solution, suspension, or powder compositions may be administered,preferably orally or nasally, from devices that deliver the formulationin an appropriate manner.

Other Pharmaceutical Compositions.

Pharmaceutical compositions may also be prepared from compositionsdescribed herein and one or more pharmaceutically acceptable excipientssuitable for sublingual, buccal, rectal, intraosseous, intraocular,intranasal, epidural, or intraspinal administration. Preparations forsuch pharmaceutical compositions are well-known in the art. See, e.g.,See, e.g., Anderson, Philip O.; Knoben, James E.; Troutman, William G,eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill, 2002;Pratt and Taylor, eds., Principles of Drug Action, Third Edition,Churchill Livingston, N.Y., 1990; Katzung, ed., Basic and ClinicalPharmacology, Ninth Edition, McGraw Hill, 20037ybg; Goodman and Gilman,eds., The Pharmacological Basis of Therapeutics, Tenth Edition, McGrawHill, 2001; Remingtons Pharmaceutical Sciences, 20th Ed., LippincottWilliams & Wilkins, 2000; Martindale, The Extra Pharmacopoeia,Thirty-Second Edition (The Pharmaceutical Press, London, 1999); all ofwhich are incorporated by reference herein in their entirety.

Administration of the compounds or pharmaceutical composition of thepresent invention can be effected by any method that enables delivery ofthe compounds to the site of action. These methods include oral routes,intraduodenal routes, parenteral injection (including intravenous,intraarterial, subcutaneous, intramuscular, intravascular,intraperitoneal or infusion), topical (e.g., transdermal application),rectal administration, via local delivery by catheter or stent orthrough inhalation. Compounds can also a be administered intraadiposallyor intrathecally.

The amount of the compound administered will be dependent on the mammalbeing treated, the severity of the disorder or condition, the rate ofadministration, the disposition of the compound and the discretion ofthe prescribing physician. However, an effective dosage is in the rangeof about 0.001 to about 100 mg per kg body weight per day, preferablyabout 1 to about 35 mg/kg/day, in single or divided doses. For a 70 kghuman, this would amount to about 0.05 to 7 g/day, preferably about 0.05to about 2.5 g/day. In some instances, dosage levels below the lowerlimit of the aforesaid range may be more than adequate, while in othercases still larger doses may be employed without causing any harmfulside effect, e.g., by dividing such larger doses into several smalldoses for administration throughout the day.

In some embodiments, a compound of the invention is administered in asingle dose. Typically, such administration will be by injection, e.g.,intravenous injection, in order to introduce the agent quickly. However,other routes may be used as appropriate. A single dose of a compound ofthe invention may also be used for treatment of an acute condition.

In some embodiments, a compound of the invention is administered inmultiple doses. Dosing may be about once, twice, three times, fourtimes, five times, six times, or more than six times per day. Dosing maybe about once a month, once every two weeks, once a week, or once everyother day. In another embodiment a compound of the invention and anotheragent are administered together about once per day to about 6 times perday. In another embodiment the administration of a compound of theinvention and an agent continues for less than about 7 days. In yetanother embodiment the administration continues for more than about 6,10, 14, 28 days, two months, six months, or one year. In some cases,continuous dosing is achieved and maintained as long as necessary.

Administration of the agents of the invention may continue as long asnecessary. In some embodiments, an agent of the invention isadministered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In someembodiments, an agent of the invention is administered for less than 28,14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, an agent of theinvention is administered chronically on an ongoing basis, e.g., for thetreatment of chronic effects.

An effective amount of a compound of the invention may be administeredin either single or multiple doses by any of the accepted modes ofadministration of agents having similar utilities, including rectal,buccal, intranasal and transdermal routes, by intra-arterial injection,intravenously, intraperitoneally, parenterally, intramuscularly,subcutaneously, orally, topically, or as an inhalant.

The compositions of the invention may also be delivered via animpregnated or coated device such as a stent, for example, or anartery-inserted cylindrical polymer. Such a method of administrationmay, for example, aid in the prevention or amelioration of restenosisfollowing procedures such as balloon angioplasty. Without being bound bytheory, compounds of the invention may slow or inhibit the migration andproliferation of smooth muscle cells in the arterial wall whichcontribute to restenosis. A compound of the invention may beadministered, for example, by local delivery from the struts of a stent,from a stent graft, from grafts, or from the cover or sheath of a stent.In some embodiments, a compound of the invention is admixed with amatrix. Such a matrix may be a polymeric matrix, and may serve to bondthe compound to the stent. Polymeric matrices suitable for such use,include, for example, lactone-based polyesters or copolyesters such aspolylactide, polycaprolactonglycolide, polyorthoesters, polyanhydrides,polyaminoacids, polysaccharides, polyphosphazenes, poly (ether-ester)copolymers (e.g., PEO-PLLA); polydimethylsiloxane,poly(ethylene-vinylacetate), acrylate-based polymers or copolymers(e.g., polyhydroxyethyl methylmethacrylate, polyvinyl pyrrolidinone),fluorinated polymers such as polytetrafluoroethylene and celluloseesters. Suitable matrices may be nondegrading or may degrade with time,releasing the compound or compounds. Compounds of the invention may beapplied to the surface of the stent by various methods such as dip/spincoating, spray coating, dip-coating, and/or brush-coating. The compoundsmay be applied in a solvent and the solvent may be allowed to evaporate,thus forming a layer of compound onto the stent. Alternatively, thecompound may be located in the body of the stent or graft, for examplein microchannels or micropores. When implanted, the compound diffusesout of the body of the stent to contact the arterial wall. Such stentsmay be prepared by dipping a stent manufactured to contain suchmicropores or microchannels into a solution of the compound of theinvention in a suitable solvent, followed by evaporation of the solvent.Excess drug on the surface of the stent may be removed via an additionalbrief solvent wash. In yet other embodiments, compounds of the inventionmay be covalently linked to a stent or graft. A covalent linker may beused which degrades in vivo, leading to the release of the compound ofthe invention. Any bio-labile linkage may be used for such a purpose,such as ester, amide or anhydride linkages. Compounds of the inventionmay additionally be administered intravascularly from a balloon usedduring angioplasty. Extravascular administration of the compounds viathe pericard or via advential application of formulations of theinvention may also be performed to decrease restenosis.

A variety of stent devices which may be used as described are disclosed,for example, in the following references, all of which are herebyincorporated by reference: U.S. Pat. No. 5,451,233; U.S. Pat. No.5,040,548; U.S. Pat. No. 5,061,273; U.S. Pat. No. 5,496,346; U.S. Pat.No. 5,292,331; U.S. Pat. No. 5,674,278; U.S. Pat. No. 3,657,744; U.S.Pat. No. 4,739,762; U.S. Pat. No. 5,195,984; U.S. Pat. No. 5,292,331;U.S. Pat. No. 5,674,278; U.S. Pat. No. 5,879,382; U.S. Pat. No.6,344,053.

The compounds of the invention may be administered in dosages. It isknown in the art that due to intersubject variability in compoundpharmacokinetics, individualization of dosing regimen is necessary foroptimal therapy. Dosing for a compound of the invention may be found byroutine experimentation in light of the instant disclosure.

When a compound of the invention, is administered in a composition thatcomprises one or more agents, and the agent has a shorter half-life thanthe compound of the invention unit dose forms of the agent and thecompound of the invention may be adjusted accordingly.

The subject pharmaceutical composition may, for example, be in a formsuitable for oral administration as a tablet, capsule, pill, powder,sustained release formulations, solution, suspension, for parenteralinjection as a sterile solution, suspension or emulsion, for topicaladministration as an ointment or cream or for rectal administration as asuppository. The pharmaceutical composition may be in unit dosage formssuitable for single administration of precise dosages. Thepharmaceutical composition will include a conventional pharmaceuticalcarrier or excipient and a compound according to the invention as anactive ingredient. In addition, it may include other medicinal orpharmaceutical agents, carriers, adjuvants, etc.

Exemplary parenteral administration forms include solutions orsuspensions of active compound in sterile aqueous solutions, forexample, aqueous propylene glycol or dextrose solutions. Such dosageforms can be suitably buffered, if desired.

The invention also provides kits. The kits include a compound orcompounds of the present invention as described herein, in suitablepackaging, and written material that can include instructions for use,discussion of clinical studies, listing of side effects, and the like.Such kits may also include information, such as scientific literaturereferences, package insert materials, clinical trial results, and/orsummaries of these and the like, which indicate or establish theactivities and/or advantages of the composition, and/or which describedosing, administration, side effects, drug interactions, or otherinformation useful to the health care provider. Such information may bebased on the results of various studies, for example, studies usingexperimental animals involving in vivo models and studies based on humanclinical trials. The kit may further contain another agent. In someembodiments, the compound of the present invention and the agent areprovided as separate compositions in separate containers within the kit.In some embodiments, the compound of the present invention and the agentare provided as a single composition within a container in the kit.Suitable packaging and additional articles for use (e.g., measuring cupfor liquid preparations, foil wrapping to minimize exposure to air, andthe like) are known in the art and may be included in the kit. Kitsdescribed herein can be provided, marketed and/or promoted to healthproviders, including physicians, nurses, pharmacists, formularyofficials, and the like. Kits may also, in some embodiments, be marketeddirectly to the consumer.

The invention also provides methods of using the compounds orpharmaceutical compositions of the present invention to treat diseaseconditions, including but not limited to diseases associated withmalfunctioning of one or more types of PI3 kinase (particularly PI3kinase α), and/or mTOR. A detailed description of conditions anddisorders mediated by p110δ kinase activity is set forth in Sadu et al.,WO 01/81346, which is incorporated herein by reference in its entiretyfor all purposes.

The invention also relates to a method of treating a hyperproliferativedisorder in a mammal that comprises administering to said mammal atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrateor derivative thereof. In some embodiments, said method relates to thetreatment of cancer such as leukemia, thymus, brain, lung, squamouscell, skin, eye, retinoblastoma, intraocular melanoma, oral cavity andoropharyngeal, bladder, gastric, stomach, pancreatic, bladder, breast,cervical head, neck, renal, kidney, liver, ovarian, prostate,colorectal, esophageal, testicular, gynecological, thyroid, CNS, PNS,AIDS-related (e.g., Lymphoma and Kaposi's Sarcoma) or viral-inducedcancer. In some embodiments, the cancer is a brain glioma, glioblastoma,leukemia, Bannayan-Zonana syndrome, Cowden disease, Lhermitte-Duclosdisease, breast cancer, inflammatory breast cancer, Wilm's tumor,Ewing's sarcoma, rhabdomyosarcoma, ependymoma, medulloblastoma, sarcoma,osteosarcoma, or a giant cell tumor of the bone or thyroid. In otherembodiments, a compound of the invention is used to treat lymphoblasticT cell leukemia, chronic myelogenous leukemia, chronic lymphocyticleukemia, hairy-cell leukemia, acute lymphoblastic leukemia, acutemyelogenous leukemia, chronic neutrophilic leukemia, acute lymphoblasticT cell leukemia, plasmacytoma, immunoblastic large cell leukemia,multiple myeloma, mantle cell leukemia, megakaryoblastic leukemia, acutemegakaryocytic leukemia, promyclocytic leukemia or erythroleukemia. Instill other embodiments, the invention provides compounds for thetreatment of malignant lymphoma, Hodgkins lymphoma, Non-Hodgkinslymphoma, lymphoblastic T cell lymphoma, Burkitt's lymphoma orfollicular lymphoma. In other embodiments, the invention relates totreatment of a cancer which is neuroblastoma, bladder cancer, urothelialcancer, vulval cancer, endometrial cancer, mesothelioma, salivary glandcancer, hepatocellular cancer, nasopharangcal cancer, buccal cancer, andgastrointestinal stromal tumors.

In some embodiments, said method relates to the treatment of anon-cancerous hyperproliferative disorder such as benign hyperplasia ofthe skin (e. g., psoriasis), restenosis, or prostate (e. g., benignprostatic hypertrophy (BPH)).

The treatment methods provided herein comprise administering to thesubject a therapeutically effective amount of a compound of theinvention. In one embodiment, the present invention provides a method oftreating an inflammation disorder, including autoimmune diseases in amammal. The method comprises administering to said mammal atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrateor derivative thereof. Examples of autoimmune diseases include but arenot limited to acute disseminated encephalomyelitis (ADEM), Addison'sdisease, antiphospholipid antibody syndrome (APS), aplastic anemia,autoimmune hepatitis, cocliac disease, Crohn's disease, Diabetesmellitus (type 1), Goodpasture's syndrome, Graves' disease,Guillain-Barré syndrome (GBS), Hashimoto's disease, lupus erythematosus,multiple sclerosis, myasthenia gravis, opsoclonus myoclonus syndrome(OMS), optic neuritis, Ord's thyroiditis, oemphigus, polyarthritis,primary biliary cirrhosis, psoriasis, rheumatoid arthritis, Reiter'ssyndrome, Takayasu's arteritis, temporal arteritis (also known as “giantcell arteritis”), warm autoimmune hemolytic anemia, Wegener'sgranulomatosis, alopecia universalis, Chagas' disease, chronic fatiguesyndrome, dysautonomia, endometriosis, hidradenitis suppurativa,interstitial cystitis, neuromyotonia, sarcoidosis, scleroderma,ulcerative colitis, vitiligo, and vulvodynia. Other disorders includebone-resorption disorders, thrombosis, lung inflammation, braininfection/inflammation, meningitis and encephalitis.

In one aspect, one or more of the subject methods may be effective inameliorating symptoms associated with rheumatoid arthritis including butnot limited to a reduction in the swelling of joints, a reduction inserum anti-collagen levels, and/or a reduction in joint pathology suchas bone resorption, cartilage damage, pannus, and/or inflammation. Inanother aspect, the subject methods are effective in reducing ankleinflammation by at least about 2%, 5%, 10%, 15%, 20%, 25%, 30%, 50%,60%, or about 75% to 90%. In another aspect, the subject methods areeffective in reducing knee inflammation by at least about 2%, 5%, 10%,15%, 20%, 25%, 30%, 50%, 60%, or about 75% to 90% or more. In stillanother aspect, the subject methods are effective in reducing serumanti-type II collagen levels by at least about 10%, 12%, 15%, 20%, 24%,25%, 30%, 35%, 50%, 60%, 75%, 80%, 86%, 87%, or about 90% or more. Inanother aspect, the subject methods are effective in reducing anklehistopathology scores by about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%,60%, 75%, 80%, 90% or more. In still another aspect, the subject methodsare effective in reducing knee histopathology scores by about 5%, 10%,15%, 20%, 25%, 30%, 40%, 50%, 60%, 75%, 80%, 90% or more.

In other embodiments, the present invention provides methods of usingthe compounds or pharmaceutical compositions to treat respiratorydiseases including but not limited to diseases affecting the lobes oflung, pleural cavity, bronchial tubes, trachea, upper respiratory tract,or the nerves and muscle for breathing. For example, methods areprovided to treat obstructive pulmonary disease. Chronic obstructivepulmonary disease (COPD) is an umbrella term for a group of respiratorytract diseases that are characterized by airflow obstruction orlimitation. Conditions included in this umbrella term are: chronicbronchitis, emphysema, and bronchiectasis.

In another embodiment, the compounds described herein are used for thetreatment of asthma. Also, the compounds or pharmaceutical compositionsdescribed herein may be used for the treatment of endotoxemia andsepsis. In one embodiment, the compounds or pharmaceutical compositionsdescribed herein are used to for the treatment of rheumatoid arthritis(RA). In yet another embodiment, the compounds or pharmaceuticalcompositions described herein is used for the treatment of contact oratopic dermatitis. Contact dermatitis includes irritant dermatitis,phototoxic dermatitis, allergic dermatitis, photoallergic dermatitis,contact urticaria, systemic contact-type dermatitis and the like.Irritant dermatitis can occur when too much of a substance is used onthe skin of when the skin is sensitive to certain substance. Atopicdermatitis, sometimes called eczema, is a kind of dermatitis, an atopicskin disease.

In other embodiments, the compounds described herein are used for thetreatment of heart conditions including atherosclerosis, hearthypertrophy, cardiac myocyte dysfunction, elevated blood pressure andvasoconstriction. The invention also relates to a method of treatingdiseases related to vasculogenesis or angiogenesis in a mammal thatcomprises administering to said mammal a therapeutically effectiveamount of a compound of the present invention, or a pharmaceuticallyacceptable salt, ester, prodrug, solvate, hydrate or derivative thereof.In some embodiments, said method is for treating a disease selected fromthe group consisting of tumor angiogenesis, chronic inflammatory diseasesuch as rheumatoid arthritis, atherosclerosis, inflammatory boweldisease, skin diseases such as psoriasis, eczema, and scleroderma,diabetes, diabetic retinopathy, retinopathy of prematurity, age-relatedmacular degeneration, hemangioma, glioma, melanoma, Kaposi's sarcoma andovarian, breast, lung, pancreatic, prostate, colon and epidermoidcancer.

Patients that can be treated with compounds of the present invention, orpharmaceutically acceptable salt, ester, prodrug, solvate, hydrate orderivative of said compounds, according to the methods of this inventioninclude, for example, patients that have been diagnosed as havingpsoriasis; restenosis; atherosclerosis; BPH; breast cancer such as aductal carcinoma in duct tissue in a mammary gland, medullarycarcinomas, colloid carcinomas, tubular carcinomas, and inflammatorybreast cancer, ovarian cancer, including epithelial ovarian tumors suchas adenocarcinoma in the ovary and an adenocarcinoma that has migratedfrom the ovary into the abdominal cavity; uterine cancer; cervicalcancer such as adenocarcinoma in the cervix epithelial includingsquamous cell carcinoma and adenocarcinomas; prostate cancer, such as aprostate cancer selected from the following: an adenocarcinoma or anadenocarcinoma that has migrated to the bone; pancreatic cancer such asepitheliod carcinoma in the pancreatic duct tissue and an adenocarcinomain a pancreatic duct; bladder cancer such as a transitional cellcarcinoma in urinary bladder, urothelial carcinomas (transitional cellcarcinomas), tumors in the urothelial cells that line the bladder,squamous cell carcinomas, adenocarcinomas, and small cell cancers;leukemia such as acute myeloid leukemia (AML), acute lymphocyticleukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairycell leukemia, myelodysplasia, myeloproliferative disorders, acutemyelogenous leukemia (AML), chronic myelogenous leukemia (CML),mastocytosis, chronic lymphocytic leukemia (CLL), multiple myeloma (MM),and myelodysplastic syndrome (MDS); bone cancer; lung cancer such asnon-small cell lung cancer (NSCLC), which is divided into squamous cellcarcinomas, adenocarcinomas, and large cell undifferentiated carcinomas,and small cell lung cancer, skin cancer such as basal cell carcinoma,melanoma, squamous cell carcinoma and actinic keratosis, which is a skincondition that sometimes develops into squamous cell carcinoma; eyeretinoblastoma; cutaneous or intraocular (eye) melanoma; primary livercancer (cancer that begins in the liver); kidney cancer; thyroid cancersuch as papillary, follicular, medullary and anaplastic; AIDS-relatedlymphoma such as diffuse large B-cell lymphoma, B-cell immunoblasticlymphoma and small non-cleaved cell lymphoma Kaposi's Sarcoma;viral-induced cancers including hepatitis B virus (HBV), hepatitis Cvirus (HCV), and hepatocellular carcinoma; human lymphotropic virus-type1 (HTLV-1) and adult T-cell leukemia/lymphoma; and human papilloma virus(HPV) and cervical cancer; central nervous system cancers (CNS) such asprimary brain tumor, which includes gliomas (astrocytoma, anaplasticastrocytoma, or glioblastoma multiforme), Oligodendroglioma, Ependymoma,Meningioma, Lymphoma, Schwannoma, and Medulloblastoma; peripheralnervous system (PNS) cancers such as acoustic neuromas and malignantperipheral nerve sheath tumor (MPNST) including neurofibromas andschwannomas, malignant fibrous cytoma, malignant fibrous histiocytoma,malignant meningioma, malignant mesothelioma, and malignant mixedMüllerian tumor, oral cavity and oropharyngeal cancer such as,hypopharyngeal cancer, laryngeal cancer, nasopharyngeal cancer, andoropharyngeal cancer; stomach cancer such as lymphomas, gastric stromaltumors, and carcinoid tumors; testicular cancer such as germ cell tumors(GCTs), which include seminomas and nonseminomas, and gonadal stromaltumors, which include Leydig cell tumors and Sertoli cell tumors; thymuscancer such as to thymomas, thymic carcinomas, Hodgkin disease,non-Hodgkin lymphomas carcinoids or carcinoid tumors; rectal cancer andcolon cancer.

The invention also relates to a method of treating diabetes in a mammalthat comprises administering to said mammal a therapeutically effectiveamount of a compound of the present invention, or a pharmaceuticallyacceptable salt, ester, prodrug, solvate, hydrate or derivative thereof.

In addition, the compounds described herein may be used to treat acne.

In addition, the compounds described herein may be used for thetreatment of arteriosclerosis, including atherosclerosis.Arteriosclerosis is a general term describing any hardening of medium orlarge arteries. Atherosclerosis is a hardening of an artery specificallydue to an atheromatous plaque.

Further the compounds described herein may be used for the treatment ofglomerulonephritis. Glomerulonephritis is a primary or secondaryautoimmune renal disease characterized by inflammation of the glomeruli.It may be asymptomatic, or present with hematuria and/or proteinuria.There are many recognized types, divided in acute, subacute or chronicglomerulonephritis. Causes are infectious (bacterial, viral or parasiticpathogens), autoimmune or paraneoplastic.

Additionally, the compounds described herein may be used for thetreatment of bursitis, lupus, acute disseminated encephalomyelitis(ADEM), addison's disease, antiphospholipid antibody syndrome (APS),aplastic anemia, autoimmune hepatitis, coeliac disease, crohn's disease,diabetes mellitus (type 1), goodpasture's syndrome, graves' disease,guillain-barré syndrome (GBS), hashimoto's disease, inflammatory boweldisease, lupus erythematosus, myasthenia gravis, opsoclonus myoclonussyndrome (OMS), optic neuritis, ord's thyroiditis, ostheoarthritis,uveoretinitis, pemphigus, polyarthritis, primary biliary cirrhosis,reiter's syndrome, takayasu's arteritis, temporal arteritis, warmautoimmune hemolytic anemia, wegener's granulomatosis, alopeciauniversalis, chagas' disease, chronic fatigue syndrome, dysautonomia,endometriosis, hidradenitis suppurativa, interstitial cystitis,neuromyotonia, sarcoidosis, scleroderma, ulcerative colitis, vitiligo,vulvodynia, appendicitis, arteritis, arthritis, blepharitis,bronchiolitis, bronchitis, cervicitis, cholangitis, cholecystitis,chorioamnionitis, colitis, conjunctivitis, cystitis, dacryoadenitis,dermatomyositis, endocarditis, endometritis, enteritis, enterocolitis,epicondylitis, epididymitis, fasciitis, fibrositis, gastritis,gastroenteritis, gingivitis, hepatitis, hidradenitis, ileitis, iritis,laryngitis, mastitis, meningitis, myelitis, myocarditis, myositis,nephritis, omphalitis, oophoritis, orchitis, osteitis, otitis,pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis,pleuritis, phlebitis, pneumonitis, proctitis, prostatitis,pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis,tendonitis, tonsillitis, uveitis, vaginitis, vasculitis, or vulvitis.

The invention also relates to a method of treating a cardiovasculardisease in a mammal that comprises administering to said mammal atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrateor derivative thereof. Examples of cardiovascular conditions include,but are not limited to, atherosclerosis, restenosis, vascular occlusion,carotid obstructive disease, or ischemic conditions.

In another aspect, the present invention provides methods of disruptingthe function of a leukocyte or disrupting a function of an osteoclast.The method includes contacting the leukocyte or the osteoclast with afunction disrupting amount of a compound of the invention.

In another aspect of the present invention, methods are provided fortreating ophthalmic disease by administering one or more of the subjectcompounds or pharmaceutical compositions to the eye of a subject.

Methods are further provided for administering the compounds of thepresent invention via eye drop, intraocular injection, intravitrealinjection, topically, or through the use of a drug eluting device,microcapsule, implant, or microfluidic device. In some cases, thecompounds of the present invention are administered with a carrier orexcipient that increases the intraocular penetrance of the compound suchas an oil and water emulsion with colloid particles having an oily coresurrounded by an interfacial film.

In some cases, the colloid particles include at least one cationic agentand at least one non-ionic surfactant such as a poloxamer, tyloxapol, apolysorbate, a polyoxyethylene castor oil derivative, a sorbitan ester,or a polyoxyl stearate. In some cases, the cationic agent is analkylamine, a tertiary alkyl amine, a quarternary ammonium compound, acationic lipid, an amino alcohol, a biguanidine salt, a cationiccompound or a mixture thereof. In some cases the cationic agent is abiguanidine salt such as chlorhexidine, polyaminopropyl biguanidine,phenformin, alkylbiguanidine, or a mixture thereof. In some cases, thequaternary ammonium compound is a benzalkonium halide, lauralkoniumhalide, cetrimide, hexadecyltrimethylammonium halide,tetradecyltrimethylammonium halide, dodecyltrimethylammonium halide,cetrimonium halide, benzethonium halide, behenalkonium halide,cetalkonium halide, cetethyldimonium halide, cetylpyridinium halide,benzododecinium halide, chlorallyl methenamine halide, myristylalkoniumhalide, stearalkonium halide or a mixture of two or more thereof. Insome cases, cationic agent is a benzalkonium chloride, lauralkoniumchloride, benzododecinium bromide, benzethenium chloride,hexadecyltrimethylammonium bromide, tetradecyltrimethylammonium bromide,dodecyltrimethylammonium bromide or a mixture of two or more thereof. Insome cases, the oil phase is mineral oil and light mineral oil, mediumchain triglycerides (MCT), coconut oil; hydrogenated oils comprisinghydrogenated cottonseed oil, hydrogenated palm oil, hydrogenate castoroil or hydrogenated soybean oil; polyoxyethylene hydrogenated castor oilderivatives comprising poluoxyl-40 hydrogenated castor oil, polyoxyl-60hydrogenated castor oil or polyoxyl-100 hydrogenated castor oil.

The invention further provides methods of modulating a PI3K and/or mTorkinase activity by contacting the kinase with an amount of an effectiveamount of compound of the invention. Modulate can be inhibiting oractivating kinase activity. In some embodiments, the invention providesmethods of inhibiting kinase activity by contacting the kinase with anamount of an effective amount of a compound of the invention insolution. In some embodiments, the invention provides methods ofinhibiting the kinase activity by contacting a cell, tissue, organ thatexpress the kinase of interest. In some embodiments, the inventionprovides methods of inhibiting kinase activity in subject including butnot limited to rodents and mammal (e.g., human) by administering intothe subject an effective amount of a compound of the invention. In someembodiments, the percentage of inhibiting exceeds 50%, 60%, 70%, 80%, or90%.

In some embodiments, the kinase is selected from the group consisting ofPI3 kinase including different isoforms such as PI3 kinase α, PI3 kinaseβ, PI3 kinase γ, PI3 kinase δ; DNA-PK; mTor, Abl, VEGFR, Ephrin receptorB4 (EphB4); TEK receptor tyrosine kinase (TIE2); FMS-related tyrosinekinase 3 (FLT-3); Platelet derived growth factor receptor (PDGFR); RET;ATM; ATR; hSmg-1; Hck; Src; Epidermal growth factor receptor (EGFR);KIT; Inulsin Receptor (IR) and IGFR.

The present invention also provides methods for combination therapies inwhich an agent known to modulate other pathways, or other components ofthe same pathway, or even overlapping sets of target enzymes are used incombination with a compound of the present invention, or apharmaceutically acceptable salt, ester, prodrug, solvate, hydrate orderivative thereof. In one aspect, such therapy includes but is notlimited to the combination of the subject compound with chemotherapeuticagents, therapeutic antibodies, and radiation treatment, to provide asynergistic or additive therapeutic effect.

For treatment of autoimmune diseases, the subject compounds orpharmaceutical compositions can be used in combination with commonlyprescribed drugs including but not limited to Enbrel®, Remicade®,Humira®, Avonex®, and Rebif®. For treatment of respiratory diseases, thesubject compounds or pharmaceutical compositions can be administered incombination with commonly prescribed drugs including but not limited toXolair®, Advair®, Singulair®, and Spiriva®.

The compounds of the invention may be formulated or administered inconjunction with other agents that act to relieve the symptoms ofinflammatory conditions such as encephalomyelitis, asthma, and the otherdiseases described herein. These agents include non-steroidalanti-inflammatory drugs (NSAIDs), e.g., acetylsalicylic acid; ibuprofen;naproxen; indomethacin; nabumetone; tolmetin; etc. Corticosteroids areused to reduce inflammation and suppress activity of the immune system.The most commonly prescribed drug of this type is Prednisone.Chloroquine (Aralen) or hydroxychloroquine (Plaquenil) may also be veryuseful in some individuals with lupus. They are most often prescribedfor skin and joint symptoms of lupus. Azathioprine (Imuran) andcyclophosphamide (Cytoxan) suppress inflammation and tend to suppressthe immune system. Other agents, e.g., methotrexate and cyclosporin areused to control the symptoms of lupus. Anticoagulants are employed toprevent blood from clotting rapidly. They range from aspirin at very lowdose which prevents platelets from sticking, to heparin/coumadin.

In another one aspect, this invention also relates to methods andpharmaceutical compositions for inhibiting abnormal cell growth in amammal which comprises an amount of a compound of the present invention,or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrateor derivative thereof, in combination with an amount of an anti-canceragent (e.g., a chemotherapeutic agent). Many chemotherapeutics arepresently known in the art and can be used in combination with thecompounds of the invention.

In some embodiments, the chemotherapeutic is selected from the groupconsisting of mitotic inhibitors, alkylating agents, anti-metabolites,intercalating antibiotics, growth factor inhibitors, cell cycleinhibitors, enzymes, topoisomerase inhibitors, biological responsemodifiers, anti-hormones, angiogenesis inhibitors, immunotherapeuticagents, proapoptotic agents, and anti-androgens. Non-limiting examplesare chemotherapeutic agents, cytotoxic agents, and non-peptide smallmolecules such as Gleevec (Imatinib Mesylate), Velcade (bortezomib),Casodex (bicalutamide), Iressa (gefitinib), and Adriamycin as well as ahost of chemotherapeutic agents. Non-limiting examples ofchemotherapeutic agents include alkylating agents such as thiotepa andcyclosphosphamide (CYTOXAN™); alkyl sulfonates such as busulfan,improsulfan and piposulfan; aziridines such as benzodopa, carboquone,meturedopa, and uredopa; ethylenimines and methylamelamines includingaltretamine, triethylenemelamine, trietylenephosphoramide,triethylenethiophosphaoramide and trimethylolomelamine; nitrogenmustards such as chlorambucil, chlornaphazine, cholophosphamide,estramustine, ifosfamide, mechlorethamine, mechlorethamine oxidehydrochloride, melphalan, novembichin, phenesterine, prednimustine,trofosfamide, uracil mustard; nitrosoureas such as carmustine,chlorozotocin, fotemustine, lomustine, nimustine, ranimustine;oxazaphosphorines; nitrosoureas; triazenes; antibiotics such asanthracyclins, actinomycins and bleomycins including aclacinomysins,actinomycin, anthramycin, azaserine, bleomycins, cactinomycin,calicheamicin, carabicin, carminomycin, carzinophilin, Casodex™,chromomycins, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin,idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin,olivomycins, peplomycin, potfiromycin, puromycin, quelamycin,rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex,zinostatin, zorubicin; anti-metabolites such as methotrexate and5-fluorouracil (5-FU); folic acid analogues such as denopterin,methotrexate, pteropterin, trimetrexate; purine analogs such asfludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacitidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine,androgens such as calusterone, dromostanolone propionate, epitiostanol,mepitiostane, testolactone; anti-adrenals such as aminoglutethimide,mitotane, trilostane; folic acid replenisher such as frolinic acid;aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine;bestrabucil; bisantrene; edatraxate; debifamine; demecolcine;diaziquone; elfomithine; elliptinium acetate; etoglucid; galliumnitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone;mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinicacid; 2-ethylhydrazide; procarbazine; PSK.R™; razoxane; sizofiran;spirogermanium; tenuazonic acid; triaziquone;2,2′,2″-trichlorotriethylamine; urethan; vindesine; dacarbazine;mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine;arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxanes, e.g.,paclitaxel (TAXOL™, Bristol-Myers Squibb Oncology, Princeton, N.J.) anddocetaxel (TAXOTERE™, Rhone-Poulenc Rorer, Antony, France); retinoicacid; esperamicins; capecitabine; gemcitabine and pharmaceuticallyacceptable salts, acids or derivatives of any of the above. Alsoincluded as suitable chemotherapeutic cell conditioners areanti-hormonal agents that act to regulate or inhibit hormone action ontumors such as anti-estrogens including for example tamoxifen(Nolvadex™), raloxifene, aromatase inhibiting 4(5)-imidazoles,4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone, andtoremifene (Fareston); and anti-androgens such as flutamide, nilutamide,bicalutamide, leuprolide, and goserelin; chlorambucil; gemcitabine;6-thioguanine; mercaptopurine; methotrexate; platinum or platinumanalogs and complexes such as cisplatin and carboplatin;anti-microtubule such as diterpenoids, including paclitaxel anddocetaxel, or Vinca alkaloids including vinblastine, vincristine,vinflunine, Vindesine, and vinorelbine; etoposide (VP-16); ifosfamide;mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine;novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate;topoisomerase I and II inhibitors including camptothecins (e.g.,camptothecin-11), topotecan, irinotecan, and epipodophyllotoxins;topoisomerase inhibitor RFS 2000; epothilone A or B;difluoromethylornithine (DMFO); histone deacetylase inhibitors;compounds which induce cell differentiation processes; gonadorelinagonists; methionine aminopeptidase inhibitors; compoundstargeting/decreasing a protein or lipid kinase activity; compounds whichtarget, decrease or inhibit the activity of a protein or lipidphosphatase; anti-androgens; bisphosphonates; biological responsemodifiers; antiproliferative antibodies; heparanase inhibitors;inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasomeinhibitors; compounds used in the treatment of hematologic malignancies;compounds which target, decrease or inhibit the activity of Flt-3; Hsp90inhibitors; temozolomide (TEMODAL®); Hsp90 inhibitors such as 17-AAG(17-allylaminogeldanamycin, NSC330507), 17-DMAG(17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545),IPI-504, CNF1010, CNF2024, CNF1010 from Conforma Therapeutics;temozolomide (TEMODAL®); kinesin spindle protein inhibitors, such asSB715992 or SB743921 from GlaxoSmithKline, or pentamidine/chlorpromazinefrom CombinatoRx; MEK inhibitors such as ARRY142886 from ArrayPioPharma, AZD6244 from AstraZeneca, PD181461 or PD0325901 from Pfizer,leucovorin, EDG binders, antileukemia compounds, ribonucleotidereductase inhibitors, S-adenosylmethionine decarboxylase inhibitors,antiproliferative antibodies or other chemotherapeutic compounds. Wheredesired, the compounds or pharmaceutical composition of the presentinvention can be used in combination with commonly prescribedanti-cancer drugs such as Herceptin®, Avastin®, Erbitux®, Rituxan®,Taxol®, Arimidex®, Taxotere®, and Velcade®. Further information oncompounds which may be used in conjunction with the compounds of theinvention is provided below.

Proteasome inhibitors include compounds which target, decrease orinhibit the activity of the proteasome. Compounds which target, decreaseor inhibit the activity of the proteasome include e.g., Bortezomid(Velcade™) and MLN 341. Matrix metalloproteinase inhibitors (“MMP”inhibitors) include, but are not limited to, collagen peptidomimetic andnonpeptidomimetic inhibitors, tetracycline derivatives, e.g.,hydroxamate peptidomimetic inhibitor batimastat and its orallybioavailable analogue marimastat (BB-2516), prinomastat (AG3340),metastat (NSC 683551) BMS-279251, BAY 12-9566, TAA211, MMI270B orAAJ996. Compounds used in the treatment of hematologic malignanciesinclude, but are not limited to, FMS-like tyrosine kinase inhibitorse.g., compounds targeting, decreasing or inhibiting the activity ofFMS-like tyrosine kinase receptors (Flt-3R); interferon,1-b-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitorse.g., compounds which target, decrease or inhibit anaplastic lymphomakinase. Compounds which target, decrease or inhibit the activity ofFMS-like tyrosine kinase receptors (Flt-3R) are especially compounds,proteins or antibodies which inhibit members of the Flt-3R receptorkinase family, e.g., PKC412, midostaurin, a staurosporine derivative,SU11248 and MLN518.

Hsp90 inhibitors include compounds such as 17-AAG(17-allylaminogeldanamycin, NSC330507), 17-DMAG(17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545),IPI-504, CNF1010, CNF2024, CNF1010 from Conforma Therapeutics;temozo-lomide (TEMODAL®); kinesin spindle protein inhibitors, such asSB715992 or SB743921 from GlaxoSmithKline, or pentamidine/chlorpromazinefrom CombinatoRx; MEK inhibitors such as ARRY142886 from ArrayPioPharma, AZD6244 from AstraZeneca, PD181461 from Pfizer, leucovorin,EDG binders, antileukemia compounds, ribonucleotide reductaseinhibitors, S-adenosylmethionine decarboxylase inhibitors,antiproliferative antibodies or other chemotherapeutic compounds.

Histone deacetylase inhibitors (or “HDAC inhibitors”) include compoundswhich inhibit a histone deacetylase and which possess antiproliferativeactivity. This includes compounds disclosed in WO 02/22577, especiallyN-hydroxy-3-[4-[[(2-hydroxyethyl)[2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propenamide,N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]amino]methyl]phenyl]-2E-2-propenamide and pharmaceutically acceptablesalts thereof. It further especially includes Suberoylanilide hydroxamicacid (SAHA).

Bisphosphonates for use in combination with the compounds of theinvention include, but are not limited to, etridonic, clodronic,tiludronic, pamidronic, alendronic, ibandronic, risedronic andzoledronic acid.

Compounds of the invention may also be used in conjunction withcompounds targeting or decreasing a protein or lipid kinase activity, aprotein or lipid phosphatase activity, or further anti-angiogeniccompounds. Such compounds include, but are not limited to, proteintyrosine kinase and/or serine and/or threonine kinase inhibitors orlipid kinase inhibitors, e.g., compounds targeting, decreasing orinhibiting the activity of the platelet-derived growth factor-receptors(PDGFR), such as compounds which target, decrease or inhibit theactivity of PDGFR, especially compounds which inhibit the PDGF receptor,e.g., a N-phenyl-2-pyrimidine-amine derivative, e.g., imatinib, SU101,SU6668 and GFR-1 11; compounds targeting, decreasing or inhibiting theactivity of the fibroblast growth factor-receptors (FGFR); compoundstargeting, decreasing or inhibiting the activity of the insulin-likegrowth factor receptor I (IGF-IR), such as compounds which target,decrease or inhibit the activity of IGF-IR, especially compounds whichinhibit the kinase activity of IGF-I receptor, such as those compoundsdisclosed in WO 02/092599 or such as OSI906, or antibodies that targetthe extracellular domain of IGF-I receptor such as CP-751871, R1507,AVE1642, IMC-A12, AMG479, MK-0646, SCH717454 or its growth factors;compounds targeting, decreasing or inhibiting the activity of the Trkreceptor tyrosine kinase family, or ephrin B4 inhibitors; compoundstargeting, decreasing or inhibiting the activity of the Ax1 receptortyrosine kinase family; compounds targeting, decreasing or inhibitingthe activity of the Ret receptor tyrosine kinase; compounds targeting,decreasing or inhibiting the activity of the Kit/SCFR receptor tyrosinekinase, e.g., imatinib; compounds targeting, decreasing or inhibitingthe activity of the C-kit receptor tyrosine kinases—(part of the PDGFRfamily), such as compounds which target, decrease or inhibit theactivity of the c-Kit receptor tyrosine kinase family, especiallycompounds which inhibit the c-Kit receptor, e.g., imatinib; compoundstargeting, decreasing or inhibiting the activity of members of the c-Ablfamily, their gene-fusion products (e.g., BCR-AbI kinase) and mutants,such as compounds which target decrease or inhibit the activity of c-Ablfamily members and their gene fusion products, e.g., aN-phenyl-2-pyrimidine-amine derivative, e.g., imatinib or nilotinib(AMN107); PD180970; AG957; NSC 680410; PD173955 from ParkeDavis; ordasatinib (BMS-354825); compounds targeting, decreasing or inhibitingthe activity of members of the protein kinase C (PKC) and Raf family ofserine/threonine kinases, members of the MEK, SRC, JAK, FAK, PDK1,PKB/Akt, and Ras/MAPK family members, and/or members of thecyclin-dependent kinase family (CDK) and are especially thosestaurosporine derivatives disclosed in U.S. Pat. No. 5,093,330, e.g.,midostaurin; examples of further compounds include e.g., UCN-01,satingol, BAY 43-9006, Bryostatin 1, Perifosine; llmofosine; RO 318220and RO 320432; GO 6976; Isis 3521; LY333531/LY379196; isochinolinecompounds such as those disclosed in WO 00/09495; FTIs; PD184352 orQAN697 (a PI3K inhibitor) or AT7519 (CDK inhibitor); compoundstargeting, decreasing or inhibiting the activity of protein-tyrosinekinase inhibitors, such as compounds which target, decrease or inhibitthe activity of protein-tyrosine kinase inhibitors include imatinibmesylate (GLEEVEC) or tyrphostin. A tyrphostin is preferably a lowmolecular weight (Mr<1500) compound, or a pharmaceutically acceptablesalt thereof, especially a compound selected from thebenzylidenemalonitrile class or the S-arylbenzenemalonirile orbisubstrate quinoline class of compounds, more especially any compoundselected from the group consisting of Tyrphostin A23/RG-50810; AG 99;Tyrphostin AG 213; Tyrphostin AG 1748; Tyrphostin AG 490; TyrphostinB44; Tyrphostin B44 (+) enantiomer, Tyrphostin AG 555; AG 494;Tyrphostin AG 556, AG957 and adaphostin(4-{[(2,5-dihydroxyphenyl)methyl]amino)-benzoic acid adamantyl ester;NSC 680410, adaphostin).

Compounds of the invention may also be used in combination withcompounds targeting, decreasing or inhibiting the activity of theepidermal growth factor family of receptor tyrosine kinases (EGFR,ErbB2, ErbB3, ErbB4 as homo- or heterodimers) and their mutants, such ascompounds which target, decrease or inhibit the activity of theepidermal growth factor receptor family are especially compounds,proteins or antibodies which inhibit members of the EGF receptortyrosine kinase family, e.g., EGF receptor. ErbB2, ErbB3 and ErbB4 orbind to EGF or EGF related ligands, and are in particular thosecompounds, proteins or monoclonal antibodies generically andspecifically disclosed in WO 97/02266, e.g., the compound of ex. 39, orin EP 0 564 409, WO 99/03854, EP 0520722. EP 0 566 226. EP 0 787 722, EP0 837 063. U.S. Pat. No. 5,747,498. WO 98/10767, WO 97/30034. WO97/49688, WO 97/38983 and, especially, WO 96/30347 (e.g., compound knownas CP 358774), WO 96/33980 (e.g., compound ZD 1839) and WO 95/03283(e.g., compound ZM105180); e.g., trastuzumab (Herceptin™), cetuximab(Erbitux™), Iressa, Tarceva, OSI-774, C1-1033, EKB-569, GW-2016, E1.1,E2.4, E2.5. E6.2. E6.4. E2.1 1. E6.3 or E7.6.3, and7H-pyrrolo-[2,3-d]pyrimidine derivatives which are disclosed in WO03/013541; and compounds targeting, decreasing or inhibiting theactivity of the c-Met receptor, such as compounds which target, decreaseor inhibit the activity of c-Met, especially compounds which inhibit thekinase activity of c-Met receptor, or antibodies that target theextracellular domain of c-Met or bind to HGF. Further anti-angiogeniccompounds include compounds having another mechanism for their activity,e.g., unrelated to protein or lipid kinase inhibition e.g., thalidomide(THALOMID) and TNP-470.

Non-receptor kinase angiogenesis inhibitors may also be useful inconjunction with the compounds of the present invention. Angiogenesis ingeneral is linked to erbB21EGFR signaling since inhibitors of erbB2 andEGFR have been shown to inhibit angiogenesis, primarily VEGF expression.Accordingly, non-receptor tyrosine kinase inhibitors may be used incombination with the compounds of the present invention. For example,anti-VEGF antibodies, which do not recognize VEGFR (the receptortyrosine kinase), but bind to the ligand; small molecule inhibitors ofintegrin (alphav beta3) that will inhibit angiogenesis; endostatin andangiostatin (non-RTK) may also prove useful in combination with thedisclosed compounds. (See Bruns C J et al (2000), Cancer Res., 60:2926-2935; Schreiber A B, Winkler M E, and Derynck R. (1986), Science,232: 1250-1253; Yen L et al. (2000), Oncogene 19: 3460-3469).

Compounds which target, decrease or inhibit the activity of a protein orlipid phosphatase include e.g., inhibitors of phosphatase 1, phosphatase2A, or CDC25, e.g., okadaic acid or a derivative thereof. Compoundswhich induce cell differentiation processes are e.g., retinoic acid, α-γ- or δ-tocopherol or α- γ- or δ-tocotrienol. Cyclooxygenase inhibitorsinclude, but are not limited to, e.g., Cox-2 inhibitors, 5-alkylsubstituted 2-arylaminophenylacetic acid and derivatives, such ascelecoxib (CELEBREX), rofecoxib (VIOXX), etoricoxib, valdecoxib or a5-alkyl-2-arylaminophenylacetic acid, e.g.,5-methyl-2-(2′-chloro-6′-fluoroanilino)phenyl acetic acid, andlumiracoxib.

Heparanase inhibitors includes compounds which target, decrease orinhibit heparin sulfate degradation, including, but not limited to,PI-88. Biological response modifiers include lymphokines andinterferons, e.g., interferon γ. Inhibitors of Ras oncogenic isoformsinclude H-Ras, K-Ras, N-Ras, and other compounds which target, decreaseor inhibit the oncogenic activity of Ras. Farnesyl transferaseinhibitors include, but are not limited to, e.g., L-744832, DK8G557 andR115777 (Zarnestra).

Telomerase inhibitors include compounds which target, decrease orinhibit the activity of telomerase. Compounds which target, decrease orinhibit the activity of telomerase are especially compounds whichinhibit the telomerase receptor, e.g., telomestatin. Methionineaminopeptidase inhibitors are, for example, compounds which target,decrease or inhibit the activity of methionine aminopeptidase. Compoundswhich target, decrease or inhibit the activity of methionineaminopeptidase are e.g., bengamide or a derivative thereof.

Antiproliferative antibodies include, but are not limited to,trastuzumab (Herceptin™), Trastuzumab-DM1, erbitux, bevacizumab(Avastin™), rituximab (Rituxan®), PRO64553 (anti-CD40) and 2C4 Antibody.By antibodies is meant e.g., intact monoclonal antibodies, polyclonalantibodies, multispecific antibodies formed from at least 2 intactantibodies, and antibodies fragments so long as they exhibit the desiredbiological activity.

For the treatment of acute myeloid leukemia (AML), compounds of theinvention can be used in combination with standard leukemia therapies,especially in combination with therapies used for the treatment of AML.In particular, compounds of the invention can be administered incombination with, e.g., farnesyl transferase inhibitors and/or otherdrugs useful for the treatment of AML, such as Daunorubicin, Adriamycin,Ara-C, VP-16, Teniposide, Mitoxantrone, Idarubicin, Carboplatinum andPKC412.

Antileukemic compound for use in combination with compounds of theinvention include, for example, Ara-C, a pyrimidine analog, which is the2′-alpha-hydroxy ribose (arabinoside) derivative of deoxycytidine. Alsoincluded is the purine analog of hypoxanthine, 6-mercaptopurine (6-MP)and fludarabine phosphate. Compounds which target, decrease or inhibitactivity of histone deacetylase (HDAC) inhibitors such as sodiumbutyrate and suberoylanilide hydroxamic acid (SAHA) inhibit the activityof the enzymes known as histone deacetylases. Specific HDAC inhibitorsinclude MS275, SAHA, FK228 (formerly FR901228), Trichostatin A andcompounds disclosed in U.S. Pat. No. 6,552,065, in particular,Λ/-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamide,or a pharmaceutically acceptable salt thereof andΛ/-hydroxy-3-[4-[(2-hydroxyethyl){2-(1/-/-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propenamide,or a pharmaceutically acceptable salt thereof, e.g., the lactate salt.

Somatostatin receptor antagonists include compounds which target, treator inhibit the somatostatin receptor such as octreotide, and SOM230(pasireotide). Tumor cell damaging approaches include approaches such asionizing radiation, e.g., ionizing radiation that occurs as eitherelectromagnetic rays (such as X-rays and gamma rays) or particles (suchas alpha and beta particles). Ionizing radiation is provided in, but notlimited to, radiation therapy and is known in the art. See Hellman,Principles of Radiation Therapy, Cancer, in Principles and Practice ofOncology, Devita et al., Eds., 4th Edition, Vol. 1, pp. 248-275 (1993).EDG binders includes immunosuppressants that modulate lymphocyterecirculation, such as FTY720.

Ribonucleotide reductase inhibitors include pyrimidine or purinenucleoside analogs including, but not limited to, fludarabine and/orcytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, cladribine,6-mercaptopurine (especially in combination with ara-C against ALL)and/or pentostatin. Ribonucleotide reductase inhibitors are e.g.,hydroxyurea or 2-hydroxy-1/-/-isoindole-1,3-dione derivatives, such asPL-1, PL-2, PL-3, PL-4, PL-5, PL-6, PL-7 or PL-8 mentioned in Nandy etal., Acta Oncologica, Vol. 33, No. 8, pp. 953-961 (1994).

S-adenosylmethionine decarboxylase inhibitors include, but are notlimited to the compounds disclosed in U.S. Pat. No. 5,461,076.

Also included are in particular those compounds, proteins or monoclonalantibodies of VEGF disclosed in WO 98/35958, e.g.,1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceuticallyacceptable salt thereof, e.g., the succinate, or in WO 00/09495, WO00/27820, WO 00/59509, WO 98/11223, WO 00/27819 and EP 0 769 947; thoseas described by Prewett et al, Cancer Res, Vol. 59, pp. 5209-5218(1999); Yuan et al., Proc Natl Acad Sci USA, Vol. 93, pp. 14765-14770(1996); Zhu et al., Cancer Res, Vol. 58, pp. 3209-3214 (1998); andMordenti et al., Toxicol Pathol, Vol. 27, No. 1, pp. 14-21 (1999); in WO00/37502 and WO 94/10202; ANGIOSTATIN, described by O'Reilly et al.,Cell, Vol. 79, pp. 315-328 (1994); ENDOSTATIN, described by O'Reilly etal., Cell, Vol. 88, pp. 277-285 (1997); anthranilic acid amides; ZD4190;ZD6474; SU5416; SU6668; bevacizumab; or anti-VEGF antibodies oranti-VEGF receptor antibodies, e.g., rhuMAb and RHUFab, VEGF aptamere.g., Macugon; FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgGIantibody, Angiozyme (RPI 4610) and Bevacizumab (Avastin™).

The compounds of the invention are also useful as co-therapeuticcompounds for use in combination with other drug substances such asanti-inflammatory, bronchodilatory or antihistamine drug substances,particularly in the treatment of obstructive or inflammatory airwaysdiseases such as those mentioned hereinbefore, for example aspotentiators of therapeutic activity of such drugs or as a means ofreducing required dosaging or potential side effects of such drugs. Acompound of the invention may be mixed with the other drug substance ina fixed pharmaceutical composition or it may be administered separately,before, simultaneously with or after the other drug substance.Accordingly the invention includes a combination of a compound of theinvention as described with an anti-inflammatory, bronchodilatory,antihistamine or anti-tussive drug substance, said compound of theinvention and said drug substance being in the same or differentpharmaceutical composition. Suitable anti-inflammatory drugs includesteroids, in particular glucocorticosteroids such as budesonide,beclamethasone dipropionate, fluticasone propionate, ciclesonide ormometasone furoate, or steroids described in WO 02/88167, WO 02/12266,WO 02/100879, WO 02/00679 (especially those of Examples 3, 11, 14, 17,19, 26, 34, 37, 39, 51, 60, 67, 72, 73, 90, 99 and 101), WO 03/035668,WO 03/048181, WO 03/062259, WO 03/064445, WO 03/072592, non-steroidalglucocorticoid receptor agonists such as those described in WO 00/00531,WO 02/10143, WO 03/082280, WO 03/082787, WO 03/104195, WO 04/005229;LTB4 antagonists such LY293111, CGS025019C, CP-195543, SC-53228, BIIL284, ONO 4057, SB 209247 and those described in U.S. Pat. No. 5,451,700;LTD4 antagonists such as montelukast and zafirlukast; PDE4 inhibitorssuch cilomilast (Ariflo® GlaxoSmithKline), Roflumilast (Byk Gulden), V-11294A (Napp), BAY19-8004 (Bayer), SCH-351591 (Schering-Plough),Arofylline (Almirall Prodesfarma), PD189659/PD168787 (Parke-Davis),AWD-12-281 (Asta Medica). CDC-801 (Celgene), SeICID™ CC-10004 (Celgene),VM554/UM565 (Vernalis), T-440 (Tanabe), KW-4490 (Kyowa Hakko Kogyo), andthose disclosed in WO 92/19594, WO 93/19749, WO 93/19750, WO 93/19751,WO 98/18796, WO 99/16766, WO 01/13953, WO 03/104204, WO 03/104205, WO03/39544, WO 04/000814, WO 04/000839, WO 04/005258, WO 04/018450, WO04/018451, WO 04/018457, WO 04/018465, WO 04/018431, WO 04/018449, WO04/018450, WO 04/018451, WO 04/018457, WO 04/018465, WO 04/019944, WO04/019945, WO 04/045607 and WO 04/037805; A2a agonists such as thosedisclosed in EP 409595A2, EP 1052264, EP 1241176, WO 94/17090, WO96/02543, WO 96/02553, WO 98/28319, WO 99/24449, WO 99/24450, WO99/24451, WO 99/38877, WO 99/41267, WO 99/67263, WO 99/67264, WO99/67265, WO 99/67266, WO 00/23457, WO 00/77018, WO 00/78774, WO01/23399, WO 01/27130, WO 01/27131, WO 01/60835, WO 01/94368, WO02/00676, WO 02/22630, WO 02/96462, WO 03/086408, WO 04/039762, WO04/039766, WO 04/045618 and WO 04/046083; A2b antagonists such as thosedescribed in WO 02/42298; and beta-2 adrenoceptor agonists such asalbuterol (salbutamol), metaproterenol, terbutaline, salmeterolfenoterol, procaterol, and especially, formoterol and pharmaceuticallyacceptable salts thereof, and compounds (in free or salt or solvateform) of formula I of WO 0075114, which document is incorporated hereinby reference, preferably compounds of the Examples thereof, as well ascompounds (in free or salt or solvate form) of formula I of WO 04/16601,and also compounds of WO 04/033412. Suitable bronchodilatory drugsinclude anticholinergic or antimuscarinic compounds, in particularipratropium bromide, oxitropium bromide, tiotropium salts and CHF 4226(Chiesi), and glycopyrrolate, but also those described in WO 01/041 18,WO 02/51841, WO 02/53564, WO 03/00840, WO 03/87094, WO 04/05285, WO02/00652, WO 03/53966, EP 424021, U.S. Pat. No. 5,171,744, U.S. Pat. No.3,714,357, WO 03/33495 and WO 04/018422.

Suitable antihistamine drug substances include cetirizine hydrochloride,acetaminophen, clemastine fumarate, promethazine, loratidine,desloratidine, diphenhydramine and fexofenadine hydrochloride,activastine, astemizole, azelastine, ebastine, epinastine, mizolastineand tefenadine as well as those disclosed in WO 03/099807, WO 04/026841and JP 2004107299.

Other useful combinations of compounds of the invention withanti-inflammatory drugs are those with antagonists of chemokinereceptors, e.g., CCR-1, CCR-2, CCR-3. CCR-4, CCR-5, CCR-6. CCR-7, CCR-8,CCR-9 and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5antagonists such as Schering-Plough antagonists SC-351 125, SCH-55700and SCH-D, Takeda antagonists such as TAK-770, and CCR-5 antagonistsdescribed in U.S. Pat. No. 6,166,037 (particularly claims 18 and 19), WO00/66558 (particularly claim 8), WO 00/66559 (particularly claim 9), WO04/018425 and WO 04/026873.

Anti-microtubule or anti-mitotic agents include phase specific agentsactive against the microtubules of tumor cells during M or the mitosisphase of the cell cycle. Examples of anti-microtubule agents include,but are not limited to, diterpenoids and vinca alkaloids. Diterpenoids,which are derived from natural sources, are phase specific anti-canceragents that operate at the G₂/M phases of the cell cycle. It is believedthat the diterpenoids stabilize the β-tubulin subunit of themicrotubules, by binding with this protein. Disassembly of the proteinappears then to be inhibited with mitosis being arrested and cell deathfollowing. Examples of diterpenoids include, but are not limited to,paclitaxel and its analog docetaxel. Paclitaxel,5β,20-epoxy-1,2α,4,7β,10β,13α-hexa-hydroxytax-11-en-9-one 4,10-diacetate2-benzoate 13-ester with (2R,3S)—N-benzoyl-3-phenylisoserine; is anatural diterpene product isolated from the Pacific yew tree Taxusbrevifolia and is commercially available as an injectable solutionTAXOL®. It is a member of the taxane family of terpenes. One mechanismfor its activity relates to paclitaxel's capacity to bind tubulin,thereby inhibiting cancer cell growth. Paclitaxel has been approved forclinical use in the treatment of refractory ovarian cancer in the UnitedStates and for the treatment of breast cancer. It is a potentialcandidate for treatment of neoplasms in the skin and head and neckcarcinomas. The compound also shows potential for the treatment ofpolycystic kidney disease, lung cancer and malaria. Treatment ofpatients with paclitaxel results in bone marrow suppression (multiplecell lineages, Ignoff, R. J. et. al, Cancer Chemotherapy Pocket Guide,1998) related to the duration of dosing above a threshold concentration(50 nM) (Kearns, C. M. et. al., Seminars in Oncology, 3(6) p. 16-23,1995). Docetaxel, (2R,3S)—N-carboxy-3-phenylisoserine, N-tert-butylester, 13-ester with5β-20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytax-1-1-en-9-one 4-acetate2-benzoate, trihydrate; is commercially available as an injectablesolution as TAXOTERE®. Docetaxel is indicated for the treatment ofbreast cancer. Docetaxel is a semisynthetic derivative of paclitaxelq.v., prepared using a natural precursor, 10-deacetyl-baccatin III,extracted from the needle of the European Yew tree. The dose limitingtoxicity of docetaxel is neutropenia.

Vinca alkaloids include phase specific anti-neoplastic agents derivedfrom the periwinkle plant. Vinca alkaloids act at the M phase (mitosis)of the cell cycle by binding specifically to tubulin. Consequently, thebound tubulin molecule is unable to polymerize into microtubules.Mitosis is believed to be arrested in metaphase with cell deathfollowing. Examples of vinca alkaloids include, but are not limited to,vinblastine, vincristine, and vinorelbine. Vinblastine,vincaleukoblastine sulfate, is commercially available as VELBAN® as aninjectable solution. Although it has possible indication as a secondline therapy of various solid tumors, it is primarily indicated in thetreatment of testicular cancer and various lymphomas including Hodgkin'sDisease, and lymphocytic and histiocytic lymphomas. Myelosuppression isthe dose limiting side effect of vinblastine. Vincristine,vincaleukoblastine, 22-oxo-, sulfate, is commercially available asONCOVIN® as an injectable solution. Vincristine is indicated for thetreatment of acute leukemias and has also found use in treatmentregimens for Hodgkin's and non-Hodgkin's malignant lymphomas. Alopeciaand neurologic effects are the most common side effect of vincristineand to a lesser extent myelosupression and gastrointestinal mucositiseffects occur. Vinorelbine,3′,4′-didehydro-4′-deoxy-C′-norvincalcukoblastine[R—(R*,R*)-2,3-dihydroxybutanedioate (1:2)(salt)], commerciallyavailable as an injectable solution of vinorelbine tartrate(NAVELBINE®), is a semisynthetic vinca alkaloid. Vinorelbine isindicated as a single agent or in combination with otherchemotherapeutic agents, such as cisplatin, in the treatment of varioussolid tumors, particularly non-small cell lung, advanced breast, andhormone refractory prostate cancers. Myelosuppression is the most commondose limiting side effect of vinorelbine.

Platinum coordination complexes include non-phase specific anti-canceragents, which interact with DNA. The platinum complexes enter tumorcells, undergo, aquation and form intra- and interstrand crosslinks withDNA causing adverse biological effects to the tumor. Examples ofplatinum coordination complexes include, but are not limited to,cisplatin and carboplatin. Cisplatin, cis-diamminedichloroplatinum, iscommercially available as PLATINOL® as an injectable solution. Cisplatinis primarily indicated in the treatment of metastatic testicular andovarian cancer and advanced bladder cancer. The primary dose limitingside effects of cisplatin are nephrotoxicity, which may be controlled byhydration and diuresis, and ototoxicity. Carboplatin, platinum, diammine[1,1-cyclobutane-dicarboxylate(2-)-O,O′], is commercially available asPARAPLATIN®) as an injectable solution. Carboplatin is primarilyindicated in the first and second line treatment of advanced ovariancarcinoma. Bone marrow suppression is the dose limiting toxicity ofcarboplatin.

Alkylating agents include non-phase anti-cancer specific agents andstrong electrophiles. Typically, alkylating agents form covalentlinkages, by alkylation, to DNA through nucleophilic moieties of the DNAmolecule such as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, andimidazole groups. Such alkylation disrupts nucleic acid function leadingto cell death. Examples of alkylating agents include, but are notlimited to, nitrogen mustards such as cyclophosphamide, melphalan, andchlorambucil; alkyl sulfonates such as busulfan; nitrosoureas such ascarmustine; and triazenes such as dacarbazine. Cyclophosphamide,2-[bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine 2-oxidemonohydrate, is commercially available as an injectable solution ortablets as CYTOXAN®. Cyclophosphamide is indicated as a single agent orin combination with other chemotherapeutic agents, in the treatment ofmalignant lymphomas, multiple myeloma, and leukemias. Alopecia, nausea,vomiting and leukopenia are the most common dose limiting side effectsof cyclophosphamide. Melphalan,4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commercially availableas an injectable solution or tablets as ALKERAN®. Melphalan is indicatedfor the palliative treatment of multiple myeloma and non-resectableepithelial carcinoma of the ovary. Done marrow suppression is the mostcommon dose limiting side effect of melphalan. Chlorambucil,4-[bis(2-chloroethyl)amino]benzenebutanoic acid, is commerciallyavailable as LEUKERAN® tablets. Chlorambucil is indicated for thepalliative treatment of chronic lymphatic leukemia, and malignantlymphomas such as lymphosarcoma, giant follicular lymphoma, andHodgkin's disease. Bone marrow suppression is the most common doselimiting side effect of chlorambucil. Busulfan, 1,4-butanedioldimethanesulfonate, is commercially available as MYLERAN® TABLETS.Busulfan is indicated for the palliative treatment of chronicmyelogenous leukemia. Bone marrow suppression is the most common doselimiting side effects of busulfan. Carmustine,1,3-[bis(2-chloroethyl)-1-nitrosourea, is commercially available assingle vials of lyophilized material as BiCNU®. Carmustine is indicatedfor the palliative treatment as a single agent or in combination withother agents for brain tumors, multiple myeloma, Hodgkin's disease, andnon-Hodgkin's lymphomas. Delayed myelosuppression is the most commondose limiting side effects of carmustine. Dacarbazine,5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide, is commerciallyavailable as single vials of material as DTIC-Dome®. Dacarbazine isindicated for the treatment of metastatic malignant melanoma and incombination with other agents for the second line treatment of Hodgkin'sDisease. Nausea, vomiting, and anorexia are the most common doselimiting side effects of dacarbazine.

Antibiotic anti-neoplastics include non-phase specific agents, whichbind or intercalate with DNA. Typically, such action results in stableDNA complexes or strand breakage, which disrupts ordinary function ofthe nucleic acids leading to cell death. Examples of antibioticanti-neoplastic agents include, but are not limited to, actinomycinssuch as dactinomycin, anthrocyclins such as daunorubicin anddoxorubicin; and bleomycins. Dactinomycin, also know as Actinomycin D,is commercially available in injectable form as COSMEGEN®. Dactinomycinis indicated for the treatment of Wilm's tumor and rhabdomyosarcoma.Nausea, vomiting, and anorexia are the most common dose limiting sideeffects of dactinomycin. Daunorubicin,(8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-a-L-lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12naphthacenedione hydrochloride, is commercially available as a liposomalinjectable form as DAUNOXOME® or as an injectable as CERUBIDINE®.Daunorubicin is indicated for remission induction in the treatment ofacute nonlymphocytic leukemia and advanced HIV associated Kaposi'ssarcoma. Myelosuppression is the most common dose limiting side effectof daunorubicin. Doxorubicin,(8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-8-glycoloyl,7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12 naphthacenedionehydrochloride, is commercially available as an injectable form as RUBEX®or ADRIAMYCIN RDF®. Doxorubicin is primarily indicated for the treatmentof acute lymphoblastic leukemia and acute myeloblastic leukemia, but isalso a useful component in the treatment of some solid tumors andlymphomas. Myelosuppression is the most common dose limiting side effectof doxorubicin. Bleomycin, a mixture of cytotoxic glycopeptideantibiotics isolated from a strain of Streptomyces verticillus, iscommercially available as BLENOXANE®. Bleomycin is indicated as apalliative treatment, as a single agent or in combination with otheragents, of squamous cell carcinoma, lymphomas, and testicularcarcinomas. Pulmonary and cutaneous toxicities are the most common doselimiting side effects of bleomycin.

Topoisomerase II inhibitors include, but are not limited to,epipodophyllotoxins. Epipodophyllotoxins are phase specificanti-neoplastic agents derived from the mandrake plant.Epipodophyllotoxins typically affect cells in the S and G₂ phases of thecell cycle by forming a ternary complex with topoisomerase II and DNAcausing DNA strand breaks. The strand breaks accumulate and cell deathfollows. Examples of epipodophyllotoxins include, but are not limitedto, etoposide and teniposide. Etoposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-ethylidene-β-D-glucopyranoside], is commercially availableas an injectable solution or capsules as VePESID® and is commonly knownas VP-16. Etoposide is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of testicular andnon-small cell lung cancers. Myelosuppression is the most common sideeffect of etoposide. The incidence of leucopenia tends to be more severethan thrombocytopenia. Teniposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-thenylidene-β-D-glucopyranoside], is commercially availableas an injectable solution as VUMON® and is commonly known as VM-26.Teniposide is indicated as a single agent or in combination with otherchemotherapy agents in the treatment of acute leukemia in children.Myelosuppression is the most common dose limiting side effect ofteniposide. Teniposide can induce both leucopenia and thrombocytopenia.Other topoisomerase II inhibitors include epirubicin, idarubicin,nemorubicin, mitoxantrone, and losoxantrone.

Antimetabolite neoplastic agents include phase specific anti-neoplasticagents that act at S phase (DNA synthesis) of the cell cycle byinhibiting DNA synthesis or by inhibiting purine or pyrimidine basesynthesis and thereby limiting DNA synthesis. Consequently, S phase doesnot proceed and cell death follows. Examples of antimetaboliteanti-neoplastic agents include, but are not limited to, fluorouracil,methotrexate, cytarabine, mercaptopurine, thioguanine, and gemcitabine.5-fluorouracil, 5-fluoro-2,4-(1H,3H) pyrimidinedione, is commerciallyavailable as fluorouracil. Administration of 5-fluorouracil leads toinhibition of thymidylate synthesis and is also incorporated into bothRNA and DNA. The result typically is cell death. 5-fluorouracil isindicated as a single agent or in combination with other chemotherapyagents in the treatment of carcinomas of the breast, colon, rectum,stomach and pancreas. Myelosuppression and mucositis are dose limitingside effects of 5-fluorouracil. Other fluoropyrimidine analogs include5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridinemonophosphate. Cytarabine,4-amino-1-β-D-arabinofuranosyl-2(1H)-pyrimidinone, is commerciallyavailable as CYTOSAR-U® and is commonly known as Ara-C. It is believedthat cytarabine exhibits cell phase specificity at S-phase by inhibitingDNA chain elongation by terminal incorporation of cytarabine into thegrowing DNA chain. Cytarabine is indicated as a single agent or incombination with other chemotherapy agents in the treatment of acuteleukemia. Other cytidine analogs include 5-azacytidine and2′,2′-difluorodeoxycytidine (gemcitabine). Cytarabine inducesleucopenia, thrombocytopenia, and mucositis. Mercaptopurine,1,7-dihydro-6H-purine-6-thione monohydrate, is commercially available asPURINETHOL®. Mercaptopurine exhibits cell phase specificity at S-phaseby inhibiting DNA synthesis by an as of yet unspecified mechanism.Mercaptopurine is indicated as a single agent or in combination withother chemotherapy agents in the treatment of acute leukemia.Myelosuppression and gastrointestinal mucositis are expected sideeffects of mercaptopurine at high doses. A useful mercaptopurine analogis azathioprine. Thioguanine, 2-amino-1,7-dihydro-6H-purine-6-thione, iscommercially available as TABLOID®. Thioguanine exhibits cell phasespecificity at S-phase by inhibiting DNA synthesis by an as of yetunspecified mechanism. Thioguanine is indicated as a single agent or incombination with other chemotherapy agents in the treatment of acuteleukemia. Myelosuppression, including leucopenia, thrombocytopenia, andanemia, is the most common dose limiting side effect of thioguanineadministration. However, gastrointestinal side effects occur and can bedose limiting. Other purine analogs include pentostatin,erythrohydroxynonyladenine, fludarabine phosphate, and cladribine.Gemcitabine, 2′-deoxy-2′,2′-difluorocytidine monohydrochloride(3-isomer), is commercially available as GEMZAR®. Gemcitabine exhibitscell phase specificity at S-phase and by blocking progression of cellsthrough the G1/S boundary. Gemcitabine is indicated in combination withcisplatin in the treatment of locally advanced non-small cell lungcancer and alone in the treatment of locally advanced pancreatic cancer.Myelosuppression, including leucopenia, thrombocytopenia, and anemia, isthe most common dose limiting side effect of gemcitabine administration.Methotrexate,N-[4-[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamicacid, is commercially available as methotrexate sodium. Methotrexateexhibits cell phase effects specifically at S-phase by inhibiting DNAsynthesis, repair and/or replication through the inhibition ofdyhydrofolic acid reductase which is required for synthesis of purinenucleotides and thymidylate. Methotrexate is indicated as a single agentor in combination with other chemotherapy agents in the treatment ofchoriocarcinoma, meningeal leukemia, non-Hodgkin's lymphoma, andcarcinomas of the breast, head, neck, ovary and bladder.Myclosuppression (leucopenia, thrombocytopenia, and anemia) andmucositis are expected side effect of methotrexate administration.

Topoisomerase I inhibitors include camptothecins such as camptothecinand camptothecin derivatives. Camptothecin cytotoxic activity isbelieved to be related to its Topoisomerase I inhibitory activity.Examples of camptothecins include, but are not limited to irinotecan andtopotecan. Irinotecan HCl,(4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbonyloxy]-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dionehydrochloride, is commercially available as the injectable solutionCAMPTOSAR®. Irinotecan is a derivative of camptothecin which binds,along with its active metabolite SN-38, to the topoisomerase I-DNAcomplex. It is believed that cytotoxicity occurs as a result ofirreparable double strand breaks caused by interaction of thetopoisomerase I:DNA:irinotecan or SN-38 ternary complex with replicationenzymes. Irinotecan is indicated for treatment of metastatic cancer ofthe colon or rectum. The dose limiting side effects of irinotecan HClare myelosuppression, including neutropenia, and GI effects, includingdiarrhea. Topotecan HCl,(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3′,4′,6,7]-indolizino[1,2-b]quinoline-3,14-(4H,12H)-dione monohydrochloride, is commercially available as theinjectable solution HYCAMTIN®. Topotecan is a derivative of camptothecinwhich binds to the topoisomerase I-DNA complex and prevents religationof singles strand breaks caused by Topoisomerase I in response totorsional strain of the DNA molecule. Topotecan is indicated for secondline treatment of metastatic carcinoma of the ovary and small cell lungcancer. The dose limiting side effect of topotecan HCl ismyelosuppression, primarily neutropenia.

Hormones and hormonal analogues are useful compounds for treatingcancers in which there is a relationship between the hormone(s) andgrowth and/or lack of growth of the cancer. Examples of hormones andhormonal analogues useful in cancer treatment include, but are notlimited to, adrenocorticosteroids such as prednisone and prednisolonewhich are useful in the treatment of malignant lymphoma and acuteleukemia in children; aminoglutethimide and other aromatase inhibitorssuch as aminoglutethimide, roglethimide, pyridoglutethimide, trilostane,testolactone, ketokonazole, vorozole, fadrozole, anastrozole, letrazole,formestane, atamestane and exemestane useful in the treatment ofadrenocortical carcinoma and hormone dependent breast carcinomacontaining estrogen receptors; progestrins such as megestrol acetateuseful in the treatment of hormone dependent breast cancer andendometrial carcinoma; estrogens, androgens, and anti-androgens such asflutamide, nilutamide, bicalutamide, cyproterone acetate and5α-reductases such as finasteride and dutasteride, useful in thetreatment of prostatic carcinoma and benign prostatic hypertrophy;anti-estrogens such as fulvestrant, tamoxifen, toremifene, raloxifene,droloxifene, iodoxyfene, as well as selective estrogen receptormodulators (SERMS) such those described in U.S. Pat. Nos. 5,681,835,5,877,219, and 6,207,716, useful in the treatment of hormone dependentbreast carcinoma and other susceptible cancers; andgonadotropin-releasing hormone (GnRH) and analogues thereof whichstimulate the release of leutinizing hormone (LH) and/or folliclestimulating hormone (FSH) for the treatment prostatic carcinoma, forinstance, LHRH agonists and antagonists such as abarelix, goserelin,goserelin acetate and luprolide. SH2/SH3 domain blockers are agents thatdisrupt SH2 or SH3 domain binding in a variety of enzymes or adaptorproteins including, PI3-K p85 subunit, Src family kinases, adaptormolecules (Shc, Crk, Nck, Grb2) and Ras-GAP. SH2/SH3 domains as targetsfor anti-cancer drugs are discussed in Smithgall, T. E. (1995), Journalof Pharmacological and Toxicological Methods. 34(3) 125-32. Inhibitorsof Serine/Threonine Kinases including MAP kinase cascade blockers whichinclude blockers of Raf kinases (rafk), Mitogen or ExtracellularRegulated Kinase (MEKs), and Extracellular Regulated Kinases (ERKs); andProtein kinase C family member blockers including blockers of PKCs(alpha, beta, gamma, epsilon, mu, lambda, iota, zeta). IkB kinase family(IKKa, IKKb), PKB family kinases, akt kinase family members, and TGFbeta receptor kinases. Such Serine/Threonine kinases and inhibitorsthereof are described in Yamamoto, T., Taya, S., Kaibuchi, K., (1999),Journal of Biochemistry. 126 (5) 799-803; Brodt, P, Samani, A., andNavab, R. (2000), Biochemical Pharmacology, 60. 1101-1107; Massague, J.,Weis-Garcia, F. (1996) Cancer Surveys. 27:41-64; Philip. P. A., andHarris, A. L. (1995), Cancer Treatment and Research. 78: 3-27. Lackey.K. et al Bioorganic and Medicinal Chemistry Letters, (10), 2000,223-226; U.S. Pat. No. 6,268,391; and Martinez-Iacaci, L., et al, Int.J. Cancer (2000), 88(1), 44-52.

Also of interest for use with the compounds of the invention areMyo-inositol signaling inhibitors such as phospholipase C blockers andMyoinositol analogues. Such signal inhibitors are described in Powis,G., and Kozikowski A., (1994) New Molecular Targets for CancerChemotherapy ed., Paul Workman and David Kerr, CRC press 1994, London.

Another group of inhibitors are signal transduction pathway inhibitorssuch as inhibitors of Ras Oncogene. Such inhibitors include inhibitorsof farnesyltransferase, geranyl-geranyl transferase, and CAAX proteasesas well as anti-sense oligonucleotides, ribozymes and immunotherapy.Such inhibitors have been shown to block ras activation in cellscontaining wild type mutant ras, thereby acting as antiproliferationagents. Ras oncogene inhibition is discussed in Scharovsky, O. G.,Rozados, V. R., Gervasoni, S. I. Matar, P. (2000), Journal of BiomedicalScience. 7(4) 292-8; Ashby, M. N. (1998), Current Opinion in Lipidology.9 (2) 99-102; and BioChim. Biophys. Acta, (19899) 1423(3):19-30.

This invention further relates to a method for using the compounds orpharmaceutical composition in combination with other tumor treatmentapproaches, including surgery, ionizing radiation, photodynamic therapy,or implants, e.g., with corticosteroids, hormones, or used asradiosensitizers.

One such approach may be, for example, radiation therapy in inhibitingabnormal cell growth or treating the hyperproliferative disorder in themammal. Techniques for administering radiation therapy are known in theart, and these techniques can be used in the combination therapydescribed herein. The administration of the compound of the invention inthis combination therapy can be determined as described herein.

Radiation therapy can be administered through one of several methods, ora combination of methods, including without limitation external-beamtherapy, internal radiation therapy, implant radiation, stereotacticradiosurgery, systemic radiation therapy, radiotherapy and permanent ortemporary interstitial brachytherapy. The term “brachytherapy,” as usedherein, refers to radiation therapy delivered by a spatially confinedradioactive material inserted into the body at or near a tumor or otherproliferative tissue disease site. The term is intended withoutlimitation to include exposure to radioactive isotopes (e.g., At-211,I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, andradioactive isotopes of Lu). Suitable radiation sources for use as acell conditioner of the present invention include both solids andliquids. By way of non-limiting example, the radiation source can be aradionuclide, such as I-125, I-131, Yb-169, Ir-192 as a solid source,I-125 as a solid source, or other radionuclides that emit photons, betaparticles, gamma radiation, or other therapeutic rays. The radioactivematerial can also be a fluid made from any solution of radionuclide(s),e.g., a solution of I-125 or I-131, or a radioactive fluid can beproduced using a slurry of a suitable fluid containing small particlesof solid radionuclides, such as Au-198, Y-90. Moreover, theradionuclide(s) can be embodied in a gel or radioactive micro spheres.

Without being limited by any theory, the compounds of the presentinvention can render abnormal cells more sensitive to treatment withradiation for purposes of killing and/or inhibiting the growth of suchcells. Accordingly, this invention further relates to a method forsensitizing abnormal cells in a mammal to treatment with radiation whichcomprises administering to the mammal an amount of a compound of thepresent invention or pharmaceutically acceptable salt, ester, prodrug,solvate, hydrate or derivative thereof, which amount is effective issensitizing abnormal cells to treatment with radiation. The amount ofthe compound, salt, or solvate in this method can be determinedaccording to the means for ascertaining effective amounts of suchcompounds described herein.

Photodynamic therapy includes therapy which uses certain chemicals knownas photosensitizing compounds to treat or prevent cancers. Examples ofphotodynamic therapy include treatment with compounds, such as e.g.,VISUDYNE and porfimer sodium. Angiostatic steroids include compoundswhich block or inhibit angiogenesis, such as, e.g., anecortave,triamcinolone, hydrocortisone, 11-α-epihydrocotisol, cortexolone,17α-hydroxyprogesterone, corticosterone, desoxycorticosterone,testosterone, estrone and dexamethasone.

Implants containing corticosteroids include compounds, such as e.g.,fluocinolone and dexamethasone. Other chemotherapeutic compoundsinclude, but are not limited to, plant alkaloids, hormonal compounds andantagonists; biological response modifiers, preferably lymphokines orinterferons; antisense oligonucleotides or oligonucleotide derivatives;shRNA or siRNA; or miscellaneous compounds or compounds with other orunknown mechanism of action.

The compounds or pharmaceutical compositions of the present inventioncan be used in combination with an amount of one or more substancesselected from anti-angiogenesis agents, signal transduction inhibitors,and antiproliferative agents.

Anti-angiogenesis agents, such as MMP-2 (matrix-metalloproteinase 2)inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, and COX-11(cyclooxygenase 11) inhibitors, can be used in conjunction with acompound of the present invention and pharmaceutical compositionsdescribed herein. Examples of useful COX-II inhibitors include CELEBREX™(alecoxib), valdecoxib, and rofecoxib. Examples of useful matrixmetalloproteinase inhibitors are described in WO 96/33172 (publishedOct. 24, 1996), WO 96/27583 (published Mar. 7, 1996), European PatentApplication No. 97304971.1 (filed Jul. 8, 1997), European PatentApplication No. 99308617.2 (filed Oct. 29, 1999), WO 98/07697 (publishedFeb. 26, 1998), WO 98/03516 (published Jan. 29, 1998), WO 98/34918(published Aug. 13, 1998), WO 98/34915 (published Aug. 13, 1998), WO98/33768 (published Aug. 6, 1998), WO 98/30566 (published Jul. 16,1998), European Patent Publication 606,046 (published Jul. 13, 1994),European Patent Publication 931, 788 (published Jul. 28, 1999), WO90/05719 (published May 31, 1990), WO 99/52910 (published Oct. 21,1999), WO 99/52889 (published Oct. 21, 1999), WO 99/29667 (publishedJun. 17, 1999), PCT International Application No. PCT/IB98/01113 (filedJul. 21, 1998), European Patent Application No. 99302232.1 (filed Mar.25, 1999), Great Britain Patent Application No. 9912961.1 (filed Jun. 3,1999), U.S. Provisional Application No. 60/148,464 (filed Aug. 12,1999), U.S. Pat. No. 5,863,949 (issued Jan. 26, 1999), U.S. Pat. No.5,861,510 (issued Jan. 19, 1999), and European Patent Publication780,386 (published Jun. 25, 1997), all of which are incorporated hereinin their entireties by reference. In some embodiments. MMP-2 and MMP-9inhibitors have little or no activity inhibiting MMP-1, or selectivelyinhibit MMP-2 and/or AMP-9 relative to the othermatrix-metalloproteinases (i. e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6,MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13). Some specificexamples of MMP inhibitors useful in the present invention are AG-3340.RO 32-3555, and RS 13-0830.

The invention also relates to a method of and to a pharmaceuticalcomposition of treating a cardiovascular disease in a mammal whichcomprises an amount of a compound of the present invention, or apharmaceutically acceptable salt, ester, prodrug, solvate, hydrate orderivative thereof, or an isotopically-labeled derivative thereof, andan amount of one or more therapeutic agents use for the treatment ofcardiovascular diseases.

Exemplary agents for use in cardiovascular disease applications areanti-thrombotic agents, e.g., prostacyclin and salicylates, thrombolyticagents, e.g., streptokinase, urokinase, tissue plasminogen activator(TPA) and anisoylated plasminogen-streptokinase activator complex(APSAC), anti-platelets agents, e.g., acetylsalicylic acid (ASA) andclopidrogel, vasodilating agents, e.g., nitrates, calcium channelblocking drugs, antiproliferative agents, e.g., colchicine andalkylating agents, intercalating agents, growth modulating factors suchas interleukins, transformation growth factor-beta and congeners ofplatelet derived growth factor, monoclonal antibodies directed againstgrowth factors, anti-inflammatory agents, both steroidal andnon-steroidal and other agents that can modulate vessel tone, function,arteriosclerosis, and the healing response to vessel or organ injurypost intervention. Antibiotics can also be included in combinations orcoatings comprised by the invention. Moreover, a coating can be used toeffect therapeutic delivery focally within the vessel wall. Byincorporation of the active agent in a swellable polymer, the activeagent will be released upon swelling of the polymer.

The compounds describe herein may be formulated or administered inconjunction with liquid or solid tissue barriers also known aslubricants. Examples of tissue barriers include, but are not limited to,polysaccharides, polyglycans, seprafilm, interceed and hyaluronic acid.

Medicaments which may be administered in conjunction with the compoundsdescribed herein include any suitable drugs usefully delivered byinhalation for example, analgesics, e.g., codeine, dihydromorphine,ergotamine, fentanyl or morphine; anginal preparations, e.g., diltiazem;antiallergics, e.g., cromoglycate, ketotifen or nedocromil;anti-infectives, e.g., cephalosporins, penicillins, streptomycin,sulphonamides, tetracyclines or pentamidine; antihistamines, e.g.,methapyrilene; anti-inflammatories, e.g., beclomethasone, flunisolide,budesonide, tipredane, triamcinolone acetonide or fluticasone;antitussives, e.g., noscapine; bronchodilators, e.g., ephedrine,adrenaline, fenoterol, formoterol, isoprenaline, metaproterenol,phenylephrine, phenylpropanolamine, pirbuterol, reproterol, rimiterol,salbutamol, salmeterol, terbutalin, isoetharine, tulobuterol,orciprenaline or(−)-4-amino-3,5-dichloro-α-[[[6-[2-(2-pyridinyl)ethoxy]hexyl]-amino]methyl]benzenemethanol;diuretics, e.g., amiloride; anticholinergics e.g., ipratropium, atropineor oxitropium; hormones, e.g., cortisone, hydrocortisone orprednisolone; xanthines e.g., aminophylline, choline theophyllinate,lysine theophyllinate or theophylline; and therapeutic proteins andpeptides, e.g., insulin or glucagon. It will be clear to a personskilled in the art that, where appropriate, the medicaments may be usedin the form of salts (e.g., as alkali metal or amine salts or as acidaddition salts) or as esters (e.g., lower alkyl esters) or as solvates(e.g., hydrates) to optimize the activity and/or stability of themedicament.

Other exemplary therapeutic agents useful for a combination therapyinclude but are not limited to agents as described above, radiationtherapy, hormone antagonists, hormones and their releasing factors,thyroid and antithyroid drugs, estrogens and progestins, androgens,adrenocorticotropic hormone; adrenocortical steroids and their syntheticanalogs; inhibitors of the synthesis and actions of adrenocorticalhormones, insulin, oral hypoglycemic agents, and the pharmacology of theendocrine pancreas, agents affecting calcification and bone turnover:calcium, phosphate, parathyroid hormone, vitamin D, calcitonin, vitaminssuch as water-soluble vitamins, vitamin B complex, ascorbic acid,fat-soluble vitamins, vitamins A, K, and E, growth factors, cytokines,chemokines, muscarinic receptor agonists and antagonists;anticholinesterase agents; agents acting at the neuromuscular junctionand/or autonomic ganglia; catecholamines, sympathomimetic drugs, andadrenergic receptor agonists or antagonists; and 5-hydroxytryptamine(5-HT, serotonin) receptor agonists and antagonists.

Therapeutic agents can also include agents for pain and inflammationsuch as histamine and histamine antagonists, bradykinin and bradykininantagonists, 5-hydroxytryptamine (serotonin), lipid substances that aregenerated by biotransformation of the products of the selectivehydrolysis of membrane phospholipids, eicosanoids, prostaglandins,thromboxanes, leukotrienes, aspirin, nonsteroidal anti-inflammatoryagents, analgesic-antipyretic agents, agents that inhibit the synthesisof prostaglandins and thromboxanes, selective inhibitors of theinducible cyclooxygenase, selective inhibitors of the induciblecyclooxygenase-2, autacoids, paracrine hormones, somatostatin, gastrin,cytokines that mediate interactions involved in humoral and cellularimmune responses, lipid-derived autacoids, eicosanoids, β-adrenergicagonists, ipratropium, glucocorticoids, methylxanthines, sodium channelblockers, opioid receptor agonists, calcium channel blockers, membranestabilizers and leukotriene inhibitors.

Additional therapeutic agents contemplated herein include diuretics,vasopressin, agents affecting the renal conservation of water, rennin,angiotensin, agents useful in the treatment of myocardial ischemia,anti-hypertensive agents, angiotensin converting enzyme inhibitors.B-adrenergic receptor antagonists, agents for the treatment ofhypercholesterolemia, and agents for the treatment of dyslipidemia.

Other therapeutic agents contemplated include drugs used for control ofgastric acidity, agents for the treatment of peptic ulcers, agents forthe treatment of gastroesophageal reflux disease, prokinetic agents,antiemetics, agents used in irritable bowel syndrome, agents used fordiarrhea, agents used for constipation, agents used for inflammatorybowel disease, agents used for biliary disease, agents used forpancreatic disease. Therapeutic agents used to treat protozoaninfections, drugs used to treat Malaria, Amebiasis, Giardiasis,Trichomoniasis, Trypanosomiasis, and/or Leishmaniasis, and/or drugs usedin the chemotherapy of helminthiasis. Other therapeutic agents includeantimicrobial agents, sulfonamides, trimethoprim-sulfamethoxazolequinolones, and agents for urinary tract infections, penicillins,cephalosporins, and other, β-Lactam antibiotics, an agent comprising anaminoglycoside, protein synthesis inhibitors, drugs used in thechemotherapy of tuberculosis, mycobacterium avium complex disease, andleprosy, antifungal agents, antiviral agents including nonretroviralagents and antiretroviral agents.

Examples of therapeutic antibodies that can be combined with a subjectcompound include but are not limited to anti-receptor tyrosine kinaseantibodies (cetuximab, panitumumab, trastuzumab), anti CD20 antibodies(rituximab, tositumomab), and other antibodies such as alemtuzumab,bevacizumab, and gemtuzumab.

Moreover, therapeutic agents used for immunomodulation, such asimmunomodulators, immunosuppressive agents, tolerogens, andimmunostimulants are contemplated by the methods herein. In addition,therapeutic agents acting on the blood and the blood-forming organs,hematopoietic agents, growth factors, minerals, and vitamins,anticoagulant, thrombolytic, and antiplatelet drugs.

Further therapeutic agents that can be combined with a subject compoundmay be found in Goodman and Gilman's “The Pharmacological Basis ofTherapeutics” Tenth Edition edited by Hardman, Limbird and Gilman or thePhysician's Desk Reference, both of which are incorporated herein byreference in their entirety.

The compounds described herein can be used in combination with theagents disclosed herein or other suitable agents, depending on thecondition being treated. Hence, in some embodiments the compounds of theinvention will be co-administer with other agents as described above.When used in combination therapy, the compounds described herein may beadministered with the second agent simultaneously or separately. Thisadministration in combination can include simultaneous administration ofthe two agents in the same dosage form, simultaneous administration inseparate dosage forms, and separate administration. That is, a compounddescribed herein and any of the agents described above can be formulatedtogether in the same dosage form and administered simultaneously.Alternatively, a compound of the present invention and any of the agentsdescribed above can be simultaneously administered, wherein both theagents are present in separate formulations. In another alternative, acompound of the present invention can be administered just followed byand any of the agents described above, or vice versa. In the separateadministration protocol, a compound of the present invention and any ofthe agents described above may be administered a few minutes apart, or afew hours apart, or a few days apart.

Administration of the compounds of the present invention can be effectedby any method that enables delivery of the compounds to the site ofaction. An effective amount of a compound of the invention may beadministered in either single or multiple doses by any of the acceptedmodes of administration of agents having similar utilities, includingrectal, buccal, intranasal and transdermal routes, by intra-arterialinjection, intravenously, intraperitoneally, parenterally,intramuscularly, subcutaneously, orally, topically, as an inhalant, orvia an impregnated or coated device such as a stent, for example, or anartery-inserted cylindrical polymer.

The amount of the compound administered will be dependent on the mammalbeing treated, the severity of the disorder or condition, the rate ofadministration, the disposition of the compound and the discretion ofthe prescribing physician. However, an effective dosage is in the rangeof about 0.001 to about 100 mg per kg body weight per day, preferablyabout 1 to about 35 mg/kg/day, in single or divided doses. For a 70 kghuman, this would amount to about 0.05 to 7 g/day, preferably about 0.05to about 2.5 g/day. In some instances, dosage levels below the lowerlimit of the aforesaid range may be more than adequate, while in othercases still larger doses may be employed without causing any harmfulside effect, e.g., by dividing such larger doses into several smalldoses for administration throughout the day.

In some embodiments, a compound of the invention is administered in asingle dose. Typically, such administration will be by injection, e.g.,intravenous injection, in order to introduce the agent quickly. However,other routes may be used as appropriate. A single dose of a compound ofthe invention may also be used for treatment of an acute condition.

In some embodiments, a compound of the invention is administered inmultiple doses. Dosing may be about once, twice, three times, fourtimes, five times, six times, or more than six times per day. Dosing maybe about once a month, once every two weeks, once a week, or once everyother day. In another embodiment a compound of the invention and anotheragent are administered together about once per day to about 6 times perday. In another embodiment the administration of a compound of theinvention and an agent continues for less than about 7 days. In yetanother embodiment the administration continues for more than about 6,10, 14, 28 days, two months, six months, or one year. In some cases,continuous dosing is achieved and maintained as long as necessary.

Administration of the agents of the invention may continue as long asnecessary. In some embodiments, an agent of the invention isadministered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In someembodiments, an agent of the invention is administered for less than 28,14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, an agent of theinvention is administered chronically on an ongoing basis, e.g., for thetreatment of chronic effects.

When a compound of the invention, is administered in a composition thatcomprises one or more agents, and the agent has a shorter half-life thanthe compound of the invention unit dose forms of the agent and thecompound of the invention may be adjusted accordingly.

In some embodiments, compounds of the invention are tested to estimatepharmacokinetic properties and expected side effect profile. Variousassays are known in the art for this purpose. For example, oralavailability can be estimated during early stages of drug development byperforming a Caco-2 permeability assay. Further, oral pharmacokineticsin humans can be approximated by extrapolating from the results ofassays in mice, rats or monkey. In some embodiments, compounds of theinvention show good oral availability across multiple species oforganisms.

Other assays examine the effect of a drug candidate on liver functionand metabolism. Cytochrome P450 (CYP) proteins are the main enzymeinvolved in metabolizing drugs administered to mammalian organisms. Assuch, undesired interaction of a drug candidate can be a significantsource of adverse drug interactions. Generally, it is desirable for adrug to not interact with CYP isozymes such as CYP1A2, CYP2C9, CYP2C19,CYP2D6, or CYP3A4. In some embodiments, a compound of the inventionexhibits an IC50 of greater than 10 μM for CYP1A2, CYP2C9, CYP2C19,CYP2D6, or CYP3A4. Additionally, liver microsome and hepatocytemetabolism assays using human preparations can be used to estimate thein-vitro half life of a drug candidate.

Cardiac toxicity is also an important consideration in evaluating drugcandidates. For example, hERG is the gene coding for the Kv11.1potassium ion channel, a protein is involved in mediating repolarizingcurrent in the cardiac action potential in the heart. Inhibition of thehERG gene product by a drug candidate can lead to an increase in therisk of sudden death and is therefore an undesirable property. In someembodiments, a compound of the invention exhibits less than 10% hERGinhibition when administered at a suitable concentration.

Mutagenicity of drug candidate compounds can be assayed via an Ames testor a modified Ames test using e.g., the liver S9 system. In someembodiments, compounds of the invention show negative activity in such atest.

Other undesired interactions of a drug candidate can also be ascertainedvia a receptor panel screen. In some embodiments, no significantinteractions are detected for compounds of the invention.

The examples and preparations provided below further illustrate andexemplify the compounds of the present invention and methods ofpreparing such compounds. It is to be understood that the scope of thepresent invention is not limited in any way by the scope of thefollowing examples and preparations. In the following examples moleculeswith a single chiral center, unless otherwise noted, exist as a racemicmixture. Those molecules with two or more chiral centers, unlessotherwise noted, exist as a racemic mixture of diastereomers. Singleenantiomers/diastereomers may be obtained by methods known to thoseskilled in the art.

EXAMPLES Example 1: Synthesis of5-(3-(pyridin-4-yl)imidazo[1,2-a]pyridin-6-yl)benzo[d]oxazol-2-amine

The scheme above describes the synthesis of a compound of the invention.A substituted pyridine such as compound 1-1 is reacted with2-chloroacetaldehyde, resulting in the halogenated imidazopyridinecompound 1-2. Compound 1-2 is coupled to a benzoxazolyl boronic acidester in Suzuki conditions to produce compound 1-4. Furtherderivatization of compound 1-4 using, for example, NBS, DMF results inhalogenation of the imidazopyridine moiety, which is then furtherreacted in an additional Suzuki coupling using pyridine boronic acid toresult in compound 1-6.

Example 2: Synthesis of2-(4-(8-(2-aminobenzo[d]ozazol-5-yl)-3-methyl-2-ozo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)phenyl)-2-methylpropanenitrile

The above reaction scheme illustrates the synthesis of a compound of theinvention 2-13. Methylation of starting material 2-1 yields compound2-2, which is subsequently reduced to the amine 2-3. In a separatereaction, compound 2-4 is converted to a salt, such as an HCl salt,which is then reacted, for example, with 2-nitrovinyl-hydroxylamine toyield compound 2-6. Further cyclization yields compound 2-7.Halogenation with a reagent such as POCl₃ results in compound 2-8, whichcan be coupled with intermediate 2-3 to yield 2-9. The nitro moiety of2-9 is subsequently reduced to an amine, and a further reaction with4-nitrophenyl carbonochloridate results in the heterocycle 2-11. Thedesired compound 2-13 is then prepared by coupling to the benzoxazolylboronate 2-12, for example in a Suzuki coupling.

Example 3: Synthesis of6-(4-(pyridin-4-yl)quinolin-6-yl)benzo[d]thiazol-2-amine

The bicyclic compound 3-2 is prepared from bromoaniline 3-1 usingdiethyl 2-(ethoxymethylene)malonate or a similar reagent. Deprotectionand removal of the carboxylic acid, followed by halogenation using areagent such as phosphorus oxychloride yields compound 3-5.Derivatization with pyridine boronate in Suzuki coupling conditionsyields 3-6, which is reacted in a second Suzuki reaction with abenzothiazolyl boronate to yield compound 3-7. Subsequent heating inhydrochloric acid in a solvent such as methanol results in removal of anacetyl group.

Example 4: IC50 Values for Selected Compounds

TABLE 2 In Vitro IC₅₀ data for selected compounds of the invention. Thefollowing symbols are used: + (greater than 10 microMolar), ++ (lessthan 10 microMolar), +++ (less than 1 microMolar), and ++++ (less than100 nM). mTOR C PI3K α PI3K β PI3K δ PC3 T47D IC₅₀ IC₅₀ IC₅₀ IC₅₀ PI3K γproliferation proliferation Mass Structure (nM) (nM) (nM) (nM) IC₅₀ (nM)(nM)* (nM) Characterization  1

+++ ++++ +++ ++++ ++++ +++ +++ Calcd: 388.1 Found: 389.0 [M + H]⁺  2

+ +++ + + +++ ++ Calcd: 396.10 Found: 397.0  [M + H]⁺  3

+++ ++++ + +++ +++ Calcd: 354.09 Found: 355.0  [M + H]⁺  4

+++ ++++ ++++ ++++ ++++ ++++ Calcd: 338.12 Found: 339.0  [M + H]⁺  5

+++ +++ + ++ +++ Calcd: 348.14 Found: 349.0  [M + H]⁺  6

++ ++++ ++ +++ ++++ ++ +++ Calcd: 327.11 Found: 328.0  [M + H]⁺  7

++++ ++++ ++++ ++++ ++++ ++++ Calcd: 474.18 Found: 475.0  M + H]⁺  8

+ ++ + +++ Calcd: 349.13 Found: 350.0  [M + H]⁺  9

+ +++ ++ +++ Calcd: 349.13 Found: 350.0  [M + H]⁺  10

+++ ++++ +++ ++++ +++ +++ Calcd: 328.11 Found: 329.0  [M + H]⁺  11

+ ++++ ++ +++ + Calcd: 326.12 Found: 327.0  [M + H]⁺  12

+ + + + Calcd: 354.09 Found: 355.0  [M + H]⁺  13

++++ +++ +++ +++ Calcd: 354.09 Found: 355.0  [M + H]⁺  14

++ + + ++ Calcd: 344.09 Found: 345.0  [M + H]⁺  15

++++ ++++ ++++ +++ +++ Calcd: 338.12 Found: 339.0  [M + H]⁺  16

++++ ++ ++ +++ +++ Calcd: 338.12 Found: 339.0  [M + H]⁺  17

++++ +++ +++ ++++ Calcd: 354.09 Found: 355.0  [M + H]⁺  18

++ + + ++ Calcd: 338.12 Found: 339.0  [M + H]⁺  19

+ + + + Calcd: 379.14 Found: 380.0  [M + H]⁺  20

++++ ++++ ++++ Calcd: 327.11 Found: 328.0  [M + H]⁺  21

+++ ++ ++ ++ Calcd: 359.17 Found: 360.0  [M + H]⁺  22

+++ ++++ ++++ ++++ ++++ Calcd: 347.14 Found: 348.0  [M + H]⁺  23

++++ +++ ++ ++++ Calcd: 360.17 Found: 361.0  [M + H]⁺  24

++++ +++ ++++ ++++ Calcd: 361.15 Found: 362.0  [M + H]⁺  25

++ + + ++  26

+++ ++++ ++++ ++++ Calcd: 341.13 Found: 342.2  [M + H]⁺  27

+++ ++ ++++ Calcd: 374.17 Found: 375.2  [M + H]⁺  28

++++ +++ ++++ Calcd: 346.14 Found: 347.2  [M + H]⁺  29

++++ ++++ ++++ Calcd: 361.15 Found: 362 .0  [M + H]⁺  30

++++ ++++ ++++ Calcd: 436.20 Found: 437.2  [M + H]⁺  31

++++ ++++ ++++ Calcd: 338.12 Found: 339.2  [M + H]⁺  32

++ + ++ Calcd: 360.16 Found: 361.2  [M + H]⁺  33

++++ +++ +++ Calcd: 389.19 Found: 390.2  [M + H]⁺  34

++++ +++ ++++ Calcd: 375.7 Found: 376.0 [M + H]⁺  35

++ ++++ ++++ ++++ ++ Calcd: 374.19 Found: 375.0  [M + H]⁺  36

++ ++++ ++ + ++ Calcd: 375.17 Found: 376.0  [M + H]⁺  37

++++ ++++ ++++ Calcd: 327.11 Found: 328.0  [M + H]⁺  38

++++ ++++ ++++ Calcd: 316.11 Found: 317.0  [M + H]⁺  39

++++ ++++ ++++ Calcd: 314.12 Found: 315.0  [M + H]⁺  40

++ ++ + ++ Calcd: 250.09 Found: 251.0  [M + H]⁺  41

++ ++++ +++ +++ ++ Calcd: 330.12 Found: 331.0  [M + H]⁺  42

+ ++++ +++ +++ ++ Calcd: 425.20 Found: 426.0  [M + H]⁺  43

++++ ++++ ++++  44

++ ++++ ++ ++ Calcd: 376.16 Found: 377.0  [M + H]⁺  45

++++ ++++ ++++ Calcd: 406.18 Found: 407.0  [M + H]⁺  46

++++ ++++ ++++ ++++ Calcd: 363.11 Found: 364.0  [M + H]⁺  47

++++ ++++ +++ +++ +++ +++ Calcd: 362.14 Found: 363.0  [M + H]⁺  48

++ + + + Calcd: 334.14 Found: 335.0  [M + H]⁺  49

++ ++++ ++ +++ ++ Calcd: 362.14 Found: 363.0  [M + H]⁺  50

+ ++++ ++ ++++ ++ Calcd: 382.12 Found: 383.0  [M + H]⁺  51

++++ +++ +++ +++ Calcd: 412.16 Found: 413.0  [M + H]⁺  52

++ ++++ ++++ ++++ Calcd: 412.16 Found: 413.0  [M + H]⁺  53

++ + + + Calcd: 335.14 Found: 336.0  [M + H]⁺  54

++ ++++ ++ ++ ++ ++ Calcd: 363.13 Found: 364.2  [M + H]⁺  55

+ ++ + + Calcd: 342.12 Found: 363.0  [M + H]⁺  56

+++ ++++ +++ + ++++ + Calcd: 342.12 Found: 343.0  [M + H]⁺  57

+++ +++ + ++ Calcd: 336.13 Found: 337.0  [M + H]⁺  58

++++ ++++ ++++ ++++ Calcd: 328.11 Found: 329.0  [M + H]⁺  59

++++ ++++ +++ ++++ Calcd: 317.10 Found: 318.0  [M + H]⁺  60

+++ ++++ ++ +++ ++++ Calcd: 402.18 Found: 403.0  [M + H]⁺  61

+ + + + Calcd: 328.11 Found: 329.0  [M + H]⁺  62

++++ ++++ ++++ ++++ ++++ Calcd: 328.11 Found: 329.0  [M + H]⁺  63

++ + + + Calcd: 362.14 Found: 363.2  [M + H]⁺  64

++ ++++ +++ + ++++ ++ Calcd: 347.14 Found: 348.0  [M + H]⁺  65

+ ++++ ++++ ++ +++ ++ Calcd: 360.17 Found: 361.2  [M + H]⁺  66

++ + + Calcd: 403.16 Found: 404.2  [M + H]⁺  67

+++ + + Calcd: 439.13 Found: 440.0  [M + H]⁺  68

++++ ++ ++ +++ Calcd: 328.11 Found: 329.0  [M + H]⁺  69

+++ ++ ++ +++ Calcd: 317.10 Found: 318.0  [M + H]⁺  70

++++ ++++ ++++ ++++ Calcd: 354.09 Found: 355.0  [M + H]⁺  71

++++ ++ +++ ++++ Calcd: 339.11 Found: 340.0  [M + H]⁺  72

+++ ++++ +++ + ++++ Calcd: 278.08 Found: 279.0  [M + H]⁺  73

+++ ++ + + Calcd: 389.19 Found: 390.2  [M + H]⁺  74

++ ++++ ++ + ++ ++ Calcd: 403.20 Found: 404.2  [M + H]⁺  75

++++ ++ + ++ Calcd: 361.15 Found: 362.0  [M + H]⁺  76

++ ++++ + + +++ ++ Calcd: 339.11 Found: 340.0  [M + H]⁺  77

+++ Calcd: 320.05 Found: 321.0  [M + H]⁺  78

++++ +++ +++ ++++ Calcd: 328.11 Found: 329.0  [M + H]⁺  79

++ + + + Calcd: 390.18 Found: 391.2  [M + H]⁺  80

+++ + + + Calcd: 377.15 Found: 378.0  [M + H]⁺  81

++++ +++ ++++ +++ Calcd: 374.19 Found: 375.2  [M + H]⁺  82

++++ ++ ++++ ++++ Calcd: 402.18 Found: 403.2  [M + H]⁺  83

++++ ++ ++++ ++++ Calcd: 377.15 Found: 378.0  [M + H]⁺  84

++++ ++++ +++ ++++ ++ Calcd: 388.20 Found: 389.2  [M + H]⁺  85

++++ ++++ ++++ ++++ Calcd: 388.16 Found: 389.0  [M + H]⁺  86

++++ +++ +++ ++++ Calcd: 424.13 Found: 425.0  [M + H]⁺  87

++++ +++ ++++ ++++ Calcd: 438.15 Found: 439.0  [M + H]⁺  88

++++ ++++ ++ +++ ++ Calcd: 346.15 Found: 347.2  [M + H]⁺  89

++++ ++ ++++ ++++ Calcd: 428.20 Found: 429.2  [M + H]⁺  90

++++ +++ ++ +++ +++ Calcd: 452.16 Found: 453.0  [M + H]⁺  91

+ + + + Calcd: 390.18 Found: 391.0  [M + H]⁺  92

+ + + + Calcd: 377.15 Found: 378.0  [M + H]⁺  93

++ +++ + + +++ Calcd: 286.09 Found: 287.0  [M + H]⁺  94

++++ +++ ++ ++++ Calcd: 342.12 Found: 343.0  [M + H]⁺  95

++++ ++ ++++ ++++ Calcd: 438.15 Found: 439.0  [M + H]⁺  96

++++ ++ +++ ++++ +++ Calcd: 361.15 Found: 362.0  [M + H]⁺  97

++ + + + Calcd: 374.14 Found: 375.0  [M + H]⁺  98

++++ + + ++++ Calcd: 395.11 Found: 396.0  [M + H]⁺  99

++++ ++ +++ +++ ++ Calcd: 361.15 Found: 362.2  [M + H]⁺ 100

++++ ++++ ++ +++ +++ Calcd: 390.18 Found: 391.0  [M + H]⁺ 101

++++ ++ +++ ++ ++ Calcd: 402.22 Found: 403.0  [M + H]⁺ 102

++++ +++ +++ ++++ Calcd: 420.17 Found: 421.0  [M + H]⁺ 103

++++ +++ +++ +++ ++ Calcd: 445.22 Found: 446.0  [M + H]⁺ 104

++ + + ++ Calcd: 377.16 Found: 378.0  [M + H]⁺ 105

+++ ++ + +++ Calcd: 364.13 Found: 365.0  [M + H]⁺ 106

++ + + ++ Calcd: 391.16 Found: 392.0  [M + H]⁺ 107

+++ + + ++ Calcd: 411.10 Found: 412.0  [M + H]⁺ 108

++++ ++ + ++++ +++ Calcd: 372.13 Found: 373.2  [M + H]⁺ 109

+ + + + Calcd: 399.10 Found: 400.0  [M + H]⁺ 110

+ + + + Calcd: 412.13 Found: 413.0  [M + H]⁺ 111

++++ ++ + + Calcd: 362.15 Found: 363.0  [M + H]⁺ 112

++++ + ++ +++ +++ Calcd: 440.13 Found: 441.0  [M + H]⁺ 113

++++ + + +++ ++ Calcd: 404.16 Found: 405.2  [M + H]⁺ 114

++++ + + +++ Calcd: 363.12 Found: 364.0  [M + H]⁺ 115

++++ ++++ + ++ ++ Calcd: 376.15 Found: 377.0  [M + H]⁺ 116

++++ +++ + +++ +++ Calcd: 363.12 Found: 364.0  [M + H]⁺ 117

++++ +++ + +++ ++ Calcd: 347.14 Found: 348.2  [M + H]⁺ 118

+++ ++++ + ++ Calcd: 360.17 Found: 361.0  [M + H]⁺ 119

++ ++++ ++ ++ ++ ++ Calcd: 418.18 Found: 419.0  [M + H]⁺ 120

++++ +++ ++ + +++ Calcd: 417.19 Found: 418.0  [M + H]⁺ 121

+ ++++ ++++ ++++ ++++ +++ Calcd: 443.21 Found: 444.2  [M + H]⁺ 122

+ + + + Calcd: 364.13 Found: 365.0  [M + H]⁺ 123

++ + + + Calcd: 377.16 Found: 378.2  [M + H]⁺ 124

+++ +++ + +++ Calcd: 363.13 Found: 364.0  [M + H]⁺ 125

+++ + ++ +++ Calcd: 379.11 Found: 380.0  [M + H]⁺ 126

+++ ++ + +++ Calcd: 379.11 Found: 380.0  [M + H]⁺ 127

+++ ++++ + ++ Calcd: 376.15 Found: 377.2  [M + H]⁺ 128

++++ ++ ++++ ++++ Calcd: 375.17 Found: 376.2  [M + H]⁺ 129

+++ ++++ +++ +++ ++++ +++ Calcd: 529.16 Found: 530.0  [M + H]⁺ 130

+ + + + Calcd: 587.14 Found: 588.0  [M + H]⁺ 131

+++ +++ +++ +++ Calcd: 374.19 Found: 375.2  [M + H]⁺ 132

+++ + + ++ Calcd: 375.17 Found: 376.2  [M + H]⁺ 133

+ ++++ +++ +++ +++ ++ Calcd: 390.18 Found: 390.2  [M]⁺ 134

++++ +++ +++ ++ +++ Calcd: 417.19 Found: 418.0  [M + H]⁺ 135

+ ++++ +++ + +++ +++ Calcd: 360.13 Found: 361.0  [M + H]⁺ 136

+++ +++ + ++ Calcd: 374.19 Found: 375.2  [M + H]⁺ 137

+ ++++ +++ +++ +++ +++ Calcd: 416.20 Found: 417.2  [M + H]⁺ 138

++++ +++ ++++ +++ + Calcd: 457.22 Found: 458.2  [M + H]⁺ 139

+ ++++ +++ +++ +++ +++ Calcd: 431.21 Found: 432.2  [M + H]⁺ 140

+++ ++ + + Calcd: 390.18 Found: 391.2  [M + H]⁺ 141

+++ + + ++ Calcd: 390.18 Found: 391.2  [M + H]⁺ 142

++ + + + Calcd: 390.18 Found: 391.2  [M + H]⁺ 143

+ ++++ ++ ++++ ++++ +++ Calcd: 391.16 Found: 392.2  [M + H]⁺ 144

+++ ++ + + Calcd: 392.14 Found: 393.0  [M + H]⁺ 145

+++ ++ + ++ Calcd: 376.16 Found: 377.2  [M + H]⁺ 146

++ ++ + + Calcd: 392.14 Found: 393.0  [M + H]⁺ 147

+++ ++ ++ ++ Calcd: 418.18 Found: 419.2  [M + H]⁺ 148

++++ + + ++ ++ Calcd: 406.18 Found: 407.2  [M + H]⁺ 149

++++ ++++ ++ +++ + Calcd: 360.17 Found: 361.0  [M + H]⁺ 150

+++ + + + Calcd: 424.16 Found: 425.0  [M + H]⁺ 151

++++ ++ +++ ++ ++ Calcd: 388.20 Found: 389.0  [M + H]⁺ 152

++++ ++++ ++ +++ ++ Calcd: 374.19 Found: 375.0  [M + H]⁺ 153

++++ ++++ +++ +++ +++ Calcd: 400.20 Found: 401.0  [M + H]⁺ 154

++++ +++ ++ ++++ +++ Calcd: 403.18 Found: 404.0  [M + H]⁺ 155

++++ ++++ +++ ++++ Calcd: 388.20 Found: 389.0  [M + H]⁺ 156

+ ++++ +++ +++ +++ +++ Calcd: 443.21 Found: 444.0  [M + H]⁺ 157

++ + + + Calcd: 388.16 Found: 389.0  [M + H]⁺ 158

++ + + + Calcd: 375.13 Found: 376.0  [M + H]⁺ 159

++++ +++ ++++ +++ ++ Calcd: 390.18 Found: 391.0  [M + H]⁺ 160

++++ ++ + + ++ Calcd: 404.20 Found: 405.0  [M + H]⁺ 161

+++ ++ ++ ++ Calcd: 416.20 Found: 417.0  [M + H]⁺ 162

+++ + + ++ Calcd: 377.15 Found: 378.0  [M + H]⁺ 163

+++ ++ ++ ++ Calcd: 446.21 Found: 447.0  [M + H]⁺ 164

+++ +++ + + Calcd: 377.15 Found: 378.0  [M + H]⁺ 165

+++ ++ ++ + +++ Calcd: 404.20 Found: 405.0  [M + H]⁺ 166

+++ ++ + ++ Calcd: 404.20 Found: 405.2  [M + H]⁺ 167

++ + + + Calcd: 424.16 Found: 425.0  [M + H]⁺ 168

++++ +++ ++ + ++ Calcd: 360.17 Found: 361.0  [M + H]⁺ 169

++ ++++ +++ +++ +++ +++ Calcd: 430.18 Found: 431.0  [M + H]⁺ 170

++ ++++ +++ +++ ++ +++ Calcd: 443.21 Found: 444.2  [M + H]⁺ 171

+ ++++ +++ +++ +++ +++ Calcd: 430.21 Found: 431.0  [M + H]⁺ 172

++++ ++++ ++++ +++ Calcd: 388.20 Found: 389.0  [M + H]⁺ 173

++++ +++ ++ ++++ ++ Calcd: 361.15 Found: 362.0  [M + H]⁺ 174

++++ +++ ++ ++++ ++ Calcd: 342.12 Found: 343.0  [M + H]⁺ 175

++++ + + ++ + Calcd: 345.16 Found: 346.0  [M + H]⁺ 176

++++ +++ +++ ++++ +++ Calcd: 331.14 Found: 332.0  [M + H]⁺ 177

++++ + + +++ + Calcd: 343.12 Found: 343.0  [M]⁺ 178

++++ ++++ +++ +++ Calcd: 360.17 Found: 361.0  [M + H]⁺ 179

++++ +++ ++ ++ ++ Calcd: 360.17 Found: 361.0  [M + H]⁺ 180

+++ + + + Calcd: 338.12 Found: 339.0  [M + H]⁺ 181

++++ ++ +++ ++++ ++ Calcd: 466.18 Found: 467.0  [M + H]⁺ 182

++++ +++ +++ +++ ++ Calcd: 402.22 Found: 403.2  [M + H]⁺ 183

++++ ++ ++ ++++ ++ Calcd: 306.11 Found: 307.0  [M + H]⁺ 184

++++ ++ + +++ ++ Calcd: 322.11 Found: 323.0  [M + H]⁺ 185

+++ + + + Calcd: 353.13 Found: 354.0  [M + H]⁺ 186

++++ ++ +++ ++++ ++ Calcd: 347.14 Found: 348.0  [M + H]⁺ 187

++++ ++ +++ ++++ +++ Calcd: 333.12 Found: 334.0  [M + H]⁺ 188

++++ +++ +++ ++++ +++ Calcd: 321.12 Found: 322.0  [M + H]⁺ 189

++++ +++ +++ +++ +++ Calcd: 337.12 Found: 338.0  [M + H]⁺ 190

++++ + +++ ++++ + Calcd: 370.12 Found: 371.0  [M + H]⁺ 191

++++ ++ ++ ++ Calcd: 418.21 Found: 419.0  [M + H]⁺ 192

++++ +++ +++ +++ Calcd: 403.18 Found: 404.0  [M + H]⁺ 193

++++ +++ ++ ++++ Calcd: 404.16 Found: 405.0  [M + H]⁺ 194

++++ + + ++++ Calcd: 353.13 Found: 354.0  [M + H]⁺ 195

+++ ++ + ++++ Calcd: 339.11 Found: 340.0  [M + H]⁺ 196

+++ ++ + ++ Calcd: 361.15 Found: 362.0  [M + H]⁺ 197

++++ ++ ++ ++++ Calcd: 333.12 Found: 334.0  [M + H]⁺ 198

++++ ++ ++ +++ Calcd: 468.16 Found: 469.0  [M + H]⁺ 199

++++ ++ ++++ ++++ Calcd: 482.17 Found: 483.0  [M + H]⁺ 200

++++ ++ ++ ++++ ++ Calcd: 349.12 Found: 350.0  [M + H]⁺ 201

++++ ++ + ++ ++ Calcd: 377.15 Found: 378.0  [M + H]⁺ 202

++++ ++ ++ +++ +++ Calcd: 363.13 Found: 364.0  [M + H]⁺ 203

++++ ++++ +++ ++++ Calcd: 391.18 Found: 392.0  [M + H]⁺ 204

++++ +++ + + Calcd: 376.16 Found: 377.0  [M + H]⁺ 205

++++ + + + Calcd: 332.13 Found: 333.0  [M + H]⁺ 206

++++ + + +++ Calcd: 320.13 Found: 321.0  [M + H]⁺ 207

++++ ++++ +++ ++++ +++ Calcd: 417.19 Found: 418.0  [M + H]⁺ 208

++++ +++ +++ ++ ++ Calcd: 417.19 Found: 418.0  [M + H]⁺ 209

++++ +++ ++ ++++ ++ Calcd: 347.14 Found: 348.0  [M + H]⁺ 210

+++ + + + Calcd: 355.12 Found: 356.0  [M + H]⁺ 211

++++ ++++ ++++ ++++ +++ Calcd: 403.18 Found: 404.0  [M + H]⁺ 212

++++ +++ +++ +++ +++ Calcd: 375.18 Found: 376.0  [M + H]⁺ 213

++++ +++ +++ + +++ Calcd: 431.21 Found: 432.0  [M + H]⁺ 214

++++ ++++ ++ +++ Calcd: 374.15 Found: 375.0  [M + H]⁺ 215

++++ +++ ++ +++ Calcd: 400.16 Found: 401.0  [M + H]⁺ 216

++++ +++ ++ ++++ Calcd: 333.12 Found: 334.0  [M + H]⁺ 217

++++ +++ +++ +++ Calcd: 431.21 Found: 432.0  [M + H]⁺ 218

+++ +++ +++ ++ Calcd: 376.20 Found: 377.2  [M + H]⁺ 219

+++ +++ +++ +++ Calcd: 374.19 Found: 375.0  [M + H]⁺ 220

++++ +++ ++++ +++ Calcd: 390.22 Found: 391.2  [M + H]⁺ 221

++++ +++ ++++ ++++ Calcd: 388.20 Found: 389.0  [M + H]⁺ 222

++++ +++ ++++ +++ Calcd: 404.20 Found: 405.0  [M + H]⁺ 223

++++ + + +++ Calcd: 425.20 Found: 426.0  [M + H]⁺ 224

++ + + + Calcd: 377.19 Found: 378.0  [M + H]⁺ 225

++++ ++++ ++++ ++++ Calcd: 443.21 Found: 444.0  [M + H]⁺ 226

+++ + ++ ++ Calcd: 429.19 Found: 430.2  [M + H]⁺ 227

++++ +++ ++++ ++ Calcd: 443.24 Found: 444.0  [M + H]⁺ 228

Calcd: 347.14 Found: 348.0  [M + H]⁺ 229

+++ +++ + ++ Calcd: 328.11 Found: 329.0  [M + H]⁺ 230

++ +++ ++ +++ Calcd: 328.11 Found: 329.0  [M + H]⁺ 231

++ ++++ +++ ++++ ++++ ++ +++ Calcd: 324.07 Found: 325.0  [M + H]⁺ 232

++ +++ +++ ++ ++++ Calcd: 282.06 Found: 283.0  [M + H]⁺ 233

+ + + + ++ Calcd: 227.08 Found: 228.0  [M + H]⁺ 234

+++ ++++ +++ ++++ ++++ ++++ +++ Calcd: 307.11 Found: 308.0  [M + H]⁺ 235

++ ++ + ++ ++ Calcd: 274.05 Found: 275.0  [M + H]⁺ 236

+ + + + Calcd: 265.10 Found: 266.0  [M + H]⁺ 237

+++ ++ ++ ++ Calcd: 285.12 Found: 286.0  [M + H]⁺ 238

+++ +++ ++ +++ Calcd: 257.09 Found: 258.0  [M + H]⁺ 239

++ +++ ++ ++ Calcd: 339.09 Found: 340.0  [M + H]⁺ 240

++ +++ ++ ++++ Calcd: 266.08 Found: 267.0  [M + H]⁺ 241

++ +++ +++ ++++ Calcd: 266.08 Found: 267.0  [M + H]⁺ 242

++++ ++++ + ++++ Calcd: 324.07 Found: 325.0  [M + H]⁺ 243

+++ ++++ + ++++ Calcd: 282.06 Found: 283.0  [M + H]⁺ 244

+ + + + Calcd: 308.09 Found: 309.0  [M + H]⁺ 245

+++ ++ + +++ Calcd: 265.10 Found: 266.0  246

+ + + + Calcd: 266.08 Found: 267.0  [M + H]⁺ 247

+ ++ + ++ Calcd: 265.10 Found: 266.2  [M + H]⁺ 248

++ +++ ++ +++ ++ Calcd: 282.06 Found: 283.0  [M + H]⁺ 249

++++ ++++ +++ ++++ Calcd: 282.06 Found: 283.0  [M + H]⁺ 250

+ + + ++++ Calcd: 324.07 Found: 325.0  [M + H]⁺ 251

++++ ++++ ++++ ++++ Calcd: 324.07 Found: 325.0  [M + H]⁺ 252

+ ++ ++ ++ 253

+ ++ ++ +++ 254

+++ + + +++ Calcd: 318.11 Found: 319.0  [M + H]⁺ 255

++ ++ ++ ++ Calcd: 375.17 Found: 376.0  [M + H]⁺ 256

++ + + + Calcd: 346.14 Found: 347.0  [M + H]⁺ 257

++++ + + + ++ Calcd: 374.15 Found: 375.0  [M + H]⁺ 258

++++ ++++ + ++++ +++ Calcd: 390.14 Found: 391.0  [M + H]⁺ 259

+++ + + ++ Calcd: 390.14 Found: 391.0  [M + H]⁺ 260

+++ + + ++ Calcd: 390.14 Found: 391.0  [M + H]⁺ 261

++++ +++ + +++ ++ Calcd: 386.19 Found: 387.0  [M + H]⁺ 262

++ ++++ +++ + +++ ++ Calcd: 347.14 Found: 348.0  [M + H]⁺ 263

++++ ++ ++ ++ ++ Calcd: 402.18 Found: 403.0  [M + H]⁺ 264

++++ ++ ++ +++ +++ Calcd: 363.13 Found: 364.0  [M + H]⁺ 265

++++ ++ ++ ++++ +++ Calcd: 363.13 Found: 364.0  [M + H]⁺ 266

++++ ++ +++ +++ ++ Calcd: 418.18 Found: 419.2  [M + H]⁺ 267

++ ++++ ++ +++ +++ +++ Calcd: 443.21 Found: 444.2  [M + H]⁺ 268

++++ ++ ++ +++ +++ Calcd: 443.21 Found: 444.2  [M + H]⁺ 269

++++ +++ +++ ++ ++ Calcd: 445.22 Found: 446.2  [M + H]⁺ 270

++ ++++ +++ +++ +++ +++ Calcd: 445.22 Found: 446.2  [M + H]⁺ 271

+++ ++ ++ + Calcd: 389.19 Found: 390.2  [M + H]⁺ 272

+++ ++ + ++ Calcd: 336.12 Found: 337.0  [M + H]⁺ 273

+++ ++++ + + ++ +++ Calcd: 402.18 Found: 403.0  [M + H]⁺ 274

++ ++++ ++ ++ ++ +++ Calcd: 415.21 Found: 416.2  [M + H]⁺ 275

++++ +++ +++ +++ +++ Calcd: 403.21 Found: 404.0  [M + H]⁺ 276

++ ++++ ++ +++ +++ +++ Calcd: 469.22 Found: 470.2  [M + H]⁺ 277

++ ++++ +++ ++ +++ +++ Calcd: 374.15 Found: 375.0  [M + H]⁺ 278

++++ +++ ++ +++ +++ Calcd: 457.22 Found: 458.2  [M + H]⁺ 279

++++ ++ ++ ++ +++ Calcd: 459.24 Found: 460.2  [M + H]⁺ 280

++ + + + Calcd: 404.20 Found: 405.2  [M + H]⁺ 281

+++ + ++ + Calcd: 391.20 Found: 392.2  [M + H]⁺ 282

+ + + + + Calcd: 364.13 Found: 365.0  [M + H]⁺ 283

++++ + + +++ ++ Calcd: 457.19 Found: 458.0  [M + H]⁺ 284

++++ ++ ++ +++ ++ Calcd: 457.19 Found: 458.0  [M + H]⁺ 285

++++ ++ +++ +++ +++ Calcd: 471.24 Found: 472.2  [M + H]⁺ 286

++++ +++ +++ ++++ +++ Calcd: 443.21 Found: 444.2  [M + H]⁺ 287

++++ ++ +++ +++ ++ Calcd: 483.24 Found: 484.2  [M + H]⁺ 288

+++ +++ +++ ++ ++ Calcd: 420.19 Found: 421.2  [M + H]⁺ 289

+++ ++ + +++ Calcd: 391.16 Found: 392.0  [M + H]⁺ 290

+++ + + + Calcd: 405.18 Found: 406.2  [M + H]⁺ 291

+++ + + + Calcd: 418.21 Found: 419.0  [M + H]⁺ 292

++ + + + Calcd: 349.15 Found: 350.0  [M + H]⁺ 293

++++ +++ ++++ +++ +++ Calcd: 404.20 Found: 405.0  [M + H]⁺ 294

++++ + ++ ++++ ++ Calcd: 384.13 Found: 385.0  [M + H]⁺ 295

+++ +++ +++ +++ Calcd: 392.20 Found: 393.0  [M + H]⁺ 296

+++ ++ +++ + Calcd: 390.18 Found: 391.0  [M + H]⁺ 297

+++ + + ++ Calcd: 441.19 Found: 442.0  [M + H]⁺ 298

++ ++++ +++ ++ ++ ++ Calcd: 433.19 Found: 434.0  [M + H]⁺ 299

++++ +++ ++++ ++++ +++ Calcd: 433.19 Found: 434.0  [M + H]⁺ 300

++ ++ + + Calcd: 328.11 Found: 329.0  [M + H]⁺ 301

++ ++ + + Calcd: 328.11 Found: 329.0  [M + H]⁺ 302

+++ ++ ++++ ++ Calcd: 406.21 Found: 407.2  [M + H]⁺ 303

++ ++++ + ++ + +++ Calcd: 431.21 Found: 432.2  [M + H]⁺ 304

+++ + + + Calcd: 404.20 Found: 405.2  [M + H]⁺ 305

++ + + + Calcd: 348.13 Found: 349.0  [M + H]⁺ 306

++++ +++ +++ +++ Calcd: 419.21 Found: 420.0  [M + H]⁺ 307

+++ +++ + + Calcd: 376.15 Found: 377.0  [M + H]⁺ 308

+++ +++ + ++ Calcd: 376.15 Found: 377.0  [M + H]⁺ 309

+++ +++ + ++ Calcd: 360.17 Found: 361.0  [M + H]⁺ 310

+ + + + Calcd: 360.17 Found: 361.2  [M + H]⁺ 311

+++ + + + Calcd: 386.12 Found: 387.0  [M + H]⁺ 312

+++ ++ +++ ++++ Calcd: 288.07 Found: 289.0  [M + H]⁺ 313

++ ++++ ++ ++ ++ Calcd: 441.23 Found: 442.2  [M + H]⁺ 314

+++ ++ ++ ++ Calcd: 386.19 Found: 387.2  [M + H]⁺ 315

+++ ++++ ++ ++ ++ Calcd: 402.18 Found: 403.2  [M + H]⁺ 316

+++ ++ + ++ Calcd: 381.14 Found: 382.0  [M + H]⁺ 317

+ + + + Calcd: 363.13 Found: 364.0  [M + H]⁺ 318

+++ ++++ ++ + +++ Calcd: 383.15 Found: 384.2  [M + H]⁺ 319

++++ +++ ++ +++ Calcd: 383.15 Found: 384.2  [M + H]⁺ 320

+ ++++ ++ ++ ++ Calcd: 397.17 Found: 398.0  [M + H]⁺ 321

+ + + + Calcd: 376.15 Found: 377.0  [M + H]⁺ 322

+ ++++ +++ ++ +++ Calcd: 402.18 Found: 403.2  [M + H]⁺ 323

+ ++++ ++ ++ +++ Calcd: 429.19 Found: 430.2  [M + H]⁺ 324

+ ++++ ++ ++ +++ Calcd: 388.20 Found: 389.2  [M + H]⁺ 325

++++ +++ ++ ++++ Calcd: 415.18 Found: 416.2  [M + H]⁺ 326

+ ++++ +++ +++ +++ Calcd: 401.20 Found: 402.2  [M + H]⁺ 327

+ ++++ ++ +++ +++ Calcd: 443.21 Found: 444.2  [M + H]⁺ 328

+ ++++ ++ ++ ++ Calcd: 479.17 Found: 480.2  [M + H]⁺ 329

+ ++++ +++ +++ ++ Calcd: 429.23 Found: 430.2  [M + H]⁺ 330

+++ +++ ++ + + Calcd: 478.21 Found: 479.2  [M + H]⁺ 331

+ ++++ ++ ++ ++ Calcd: 443.24 Found: 444.2  [M + H]⁺ 332

+++ ++++ ++ +++ Calcd: 374.19 Found: 375.2  [M + H]⁺ 333

+ ++++ ++ +++ ++ +++ Calcd: 445.22 Found: 446.2  [M + H]⁺ 334

++++ ++ ++ ++ +++ Calcd: 450.15 Found: 451.0  [M + H]⁺ 335

+ ++++ ++ ++ + +++ Calcd: 455.24 Found: 456.2  [M + H]⁺ 336

+++ +++ ++ + Calcd: 374.19 Found: 375.2  [M + H]⁺ 337

++++ + + +++ Calcd: 411.18 Found: 412.2  [M + H]⁺ 338

++++ ++ ++ ++ ++ Calcd: 425.20 Found: 426.2  [M + H]⁺ 339

++++ +++ +++ +++ Calcd: 397.17 Found: 398.2  [M + H]⁺ 340

+++ + + +++ Calcd: 319.11 Found: 320.2  [M + H]⁺ 341

+++ + ++ ++ ++ Calcd: 416.20 Found: 417.2  [M + H]⁺ 342

+ + + + Calcd: 360.17 Found: 361.2  [M + H]⁺ 343

+ ++ + + Calcd: 360.17 Found: 361.2  [M + H]⁺ 344

++++ ++ +++ ++ Calcd: 425.20 Found: 426.2  [M + H]⁺ 345

++ ++++ +++ +++ +++ Calcd: 411.18 Found: 412.2  [M + H]⁺ 346

++++ ++ + +++ Calcd: 347.14 Found: 348.2  [M + H]⁺ 347

+++ ++ + +++ Calcd: 363.13 Found: 364.2  [M + H]⁺ 348

+++ +++ ++++ ++ +++ Calcd: 420.20 Found: 421.2  [M + H]⁺ 349

+ + + + Calcd: 377.15 Found: 378.2  [M + H]⁺ 350

+ ++++ ++ ++ + Calcd: 415.21 Found: 416.2  [M + H]⁺ 351

++ +++ + + Calcd: 374.19 Found: 375.0  [M + H]⁺ 352

+ + + + Calcd: 377.15 Found: 378.2  [M + H]⁺ 353

+ ++ + + + Calcd: 416.20 Found: 417.2  [M + H]⁺ 354

++ ++++ ++ ++ ++ +++ Calcd: 397.17 Found: 398.0  [M + H]⁺ 355

++++ ++ +++ ++ +++ Calcd: 415.21 Found: 416.2  [M + H]⁺ 356

++++ ++++ ++++ ++++ +++ Calcd: 327.11 Found: 328.2  [M + H]⁺ 357

+++ ++ ++ +++ Calcd: 377.15 Found: 378.2  [M + H]⁺ 358

++++ +++ +++ +++ Calcd: 436.14 Found: 437.0  [M + H]⁺ 359

++++ +++ ++ ++++ Calcd: 397.09 Found: 398.0  [M + H]⁺ 360

++++ +++ ++ ++++ Calcd: 381.12 Found: 382.0  [M + H]⁺ 361

++++ +++ ++ +++ Calcd: 420.17 Found: 421.0  [M + H]⁺ 362

+ + + + Calcd: 416.20 Found: 417.0  [M + H]⁺ 363

+ + + + 364

+++ ++ + +++ Calcd: 377.15 Found: 378.0  [M + H]⁺ 365

++ ++ ++ ++ Calcd: 416.20 Found: 417.0  [M + H]⁺ 366

++++ ++ + ++ Calcd: 360.17 Found: 361.0  [M + H]⁺ 367

+++ +++ ++ +++ Calcd: 332.14 Found: 333.0  [M + H]⁺ 368

++++ ++ +++ +++ Calcd: 361.15 Found: 362.0  [M + H]⁺ 369

+ + + + Calcd: 377.15 Found: 378.0  [M + H]⁺ 370

++ + + + Calcd: 397.09 Found: 398.0  [M + H]⁺ 371

+++ ++ + + Calcd: 436.14 Found: 437.0  [M + H]⁺ 372

+++ ++ ++ ++ Calcd: 381.12 Found: 382.2  [M + H]⁺ 373

+ ++++ ++ ++ ++ Calcd: 420.17 Found: 421.2  [M + H]⁺ 374

+++ ++++ ++ + Calcd: 374.19 Found: 375.2  [M + H]⁺ 375

+++ ++ +++ +++ Calcd: 416.20 Found: 417.2  [M + H]⁺ 376

++++ ++ ++ ++++ Calcd: 416.20 Found: 417.2  [M + H]⁺ 377

++ ++++ ++ ++ +++ Calcd: 507.21 Found: 508.2  [M + H]⁺ 378

+++ ++++ ++++ ++ ++ Calcd: 374.19 Found: 375.2  [M + H]⁺ 379

++ ++++ +++ ++ + Calcd: 429.23 Found: 430.2  [M + H]⁺ 380

+++ ++ + ++ Calcd: 376.16 Found: 377.0  [M + H]⁺ 381

++++ +++ ++ ++ Calcd: 429.23 Found: 430.2  [M + H]⁺ 382

++++ +++ ++ +++ Calcd: 374.19 Found: 375.20 [M + H]⁺ 383

+++ + + + Calcd: 416.20 Found: 417.2  [M + H]⁺ 384

+++ ++++ +++ ++ +++ Calcd: 414.14 Found: 415.2  [M + H]⁺ 385

++++ ++ + + Calcd: 359.17 Found: 360.2  [M + H]⁺ 386

++++ + + Calcd: 415.21 Found: 416.2  [M + H]⁺ 387

++++ +++ + +++ Calcd: 404.20 Found: 405.2  [M + H]⁺ 388

++++ ++++ +++ +++ +++ Calcd: 429.23 Found: 430.2  [M + H]⁺ 389

+++ + + ++ Calcd: 360.13 Found: 361.0  [M + H]⁺ 390

+++ + + ++ Calcd: 374.15 Found: 375.2  [M + H]⁺ 391

++ + + + Calcd: 388.16 Found: 389.2  [M + H]⁺ 392

+ + + + Calcd: 402.18 Found: 403.2  [M + H]⁺ 393

++ + + + Calcd: 414.18 Found: 415.2  [M + H]⁺ 394

++++ ++++ +++ ++++ Calcd: 383.15 Found: 384.2  [M + H]⁺ 395

++++ + + ++++ Calcd: 428.16 Found: 429.2  [M + H]⁺ 396

++++ ++++ + ++ Calcd: 359.17 Found: 360.2  [M + H]⁺ 397

++++ +++ +++ ++++ Calcd: 341.13 Found: 342.2  [M + H]⁺ 398

+++ ++ ++ ++ Calcd: 446.21 Found: 447.2  [M + H]⁺ 399

+ + + + Calcd: 365.13 Found: 366.2  [M + H]⁺ 400

++++ ++ + ++ Calcd: 364.13 Found: 365.2  [M + H]⁺ 401

+ + + ++ Calcd: 325.10 Found: 326.0  [M + H]⁺ 402

+ + + + Calcd: 345.07 Found: 346.0  [M + H]⁺ 403

++ + + + Calcd: 375.18 Found: 376.2  [M + H]⁺ 404

+ + + + Calcd: 417.19 Found: 418.2  [M + H]⁺ 405

+ + + + Calcd: 441.11 Found: 442.0  [M + H]⁺ 406

++ ++ + + Calcd: 377.15 Found: 378.2  [M + H]⁺ 407

+ ++ + + Calcd: 439.16 Found: 440.2  [M + H]⁺ 408

+ + + + Calcd: 460.22 Found: 461.2  [M + H]⁺ 409

++ + + + Calcd: 403.16 Found: 404.2  [M + H]⁺ 410

++ + + + Calcd: 391.13 Found: 392.0  [M + H]⁺ 411

++++ ++ ++ +++ Calcd: 436.15 Found: 437.2  [M + H]⁺ 412

++ + + + Calcd: 364.13 Found: 365.0  [M + H]⁺ 413

++++ +++ ++ +++ Calcd: 396.15 Found: 397.2  [M + H]⁺ 414

++ + + + Calcd: 353.15 Found: 354.2  [M + H]⁺ 415

++ + + + Calcd: 357.10 Found: 358.0  [M + H]⁺ 416

++++ ++ ++ +++ Calcd: 431.13 Found: 432.0  [M + H]⁺ 417

+++ ++++ ++++ ++++ ++++ Calcd: 529.12 Found: 530.0  [M + H]⁺ 418

++++ ++++ ++++ ++++ ++++ Calcd: 435.08 Found: 436.0  [M + H]⁺ 419

+++ ++++ ++++ ++++ ++++ Calcd: 533.07 Found: 534.0  [M + H]⁺ 420

++++ + +++ +++ Calcd: 493.14 Found: 494.2  [M + H]⁺ 421

++++ +++ ++++ ++++ Calcd: 511.13 Found: 512.0  [M + H]⁺ 422

++++ ++ +++ ++++ Calcd: 511.13 Found: 512.0  [M + H]⁺ 423

++++ +++ ++++ ++++ Calcd: 511.13 Found: 512.0  [M + H]⁺ 424

+++ + ++ +++ Calcd: 463.51 Found: 464.0  [M + H]⁺ 425

++++ + +++ ++++ Calcd: 499.49 Found: 500.0  [M + H]⁺ 426

+ + + + Calcd: 307.11 Found: 308.0  [M + H]⁺ 427

+ + + + Calcd: 265.27 Found: 266.0  [M + H]⁺ 428

++ ++ + +++ Calcd: 265.10 Found: 266.0  [M + H]⁺ 429

+ + + + Calcd: 385.15 Found: 386.2  [M + H]⁺ 430

++ ++ + ++ Calcd: 307.11 Found: 308.0  [M + H]⁺ 431

+ + + + Calcd: 307.11 Found: 308.0  [M + H]⁺ 432

++ +++ + ++++ Calcd: 284.07 Found: 285.0  [M + H]⁺ 433

+++ +++ ++ ++++ Calcd: 284.07 Found: 285.2  [M + H]⁺ 434

++++ ++++ ++++ ++++ Calcd: 342.06 Found: 343.0  [M + H]⁺ 435

++ ++ + ++++ Calcd: 342.06 Found: 343.0  [M + H]⁺ 436

++++ ++++ +++ ++++ Calcd: 300.05 Found: 301.1  [M + H]⁺ 437

++ +++ + +++ Calcd: 300.05 Found: 301.2  [M + H]⁺ 438

++ + + + Calcd: 381.12 Found: 382.2  [M + H]⁺ 439

++ + + + Calcd: 381.12 Found: 382.2  [M + H]⁺ 440

+ + + + Calcd: 462.14 Found: 463.0  [M + H]⁺ 441

+ + + + 442

+ + + + Calcd: 400.12 Found: 401.0  [M + H]⁺ 443

+ + + + Calcd: 400.12 Found: 401.2  [M + H]⁺ 444

++ + + + Calcd: 492.15 Found: 493.0  [M + H]⁺ 445

+ + + + Calcd: 430.13 Found: 431.0  [M + H]⁺ 446

+++ + + + Calcd: 376.16 Found: 377.2  [M + H]⁺ 447

++ + + + Calcd: 463.13 Found: 464.0  [M + H]⁺ 448

+++ + + ++ Calcd: 377.40 Found: 378.2  [M + H]⁺ 449

+++ +++ + ++++ Calcd: 377.15 Found: 378.2  [M + H]⁺ 450

++ + + + Calcd: 401.12 Found: 402.2  [M + H]⁺ 451

++++ +++ ++++ ++++ +++ Calcd: 497.95 Found: 498.0  [M]⁺ 452

++++ +++ ++++ ++++ +++ Calcd: 515.08 Found: 516.0  [M + H]⁺ 453

++ + + ++ Calcd: 462.14 Found: 463.0  [M + H]⁺ 454

++ + ++ ++ Calcd: 498.12 Found: 499.0  [M + H]⁺ 455

++ + + ++ Calcd: 492.15 Found: 493.2  [M + H]⁺ 456

+ + + + Calcd: 430.13 Found: 431.0  [M + H]⁺ 457

++++ ++++ ++++ ++++ +++ Calcd: 515.08 Found: 516.0  [M + H]⁺ 458

++++ ++++ ++++ ++++ +++ Calcd: 515.08 Found: 516.0  [M + H]⁺ 459

+++ + + +++ Calcd: 496.10 Found: 497.0  [M + H]⁺ 460

++ + + + Calcd: 434.08 Found: 435.0  [M + H]⁺ 461

+++ ++ ++ +++ Calcd: 532.08 Found: 533.0  [M + H]⁺ 462

++++ ++++ ++++ ++++ +++ Calcd: 494.14 Found: 495.0  [M + H]⁺ 463

++++ ++ ++ +++ Calcd: 377.15 Found: 378.2  [M + H]⁺ 464

++++ + + +++ ++ Calcd: 377.15 Found: 378.2  [M + H]⁺ 465

++++ + + +++ Calcd: 432.19 Found: 433.2  [M + H]⁺ 466

+++ ++ ++ +++ Calcd: 432.19 Found: 433.2  [M + H]⁺ 467

++++ ++ ++ ++++ +++ Calcd: 363.13 Found: 364.0  [M + H]⁺ 468

++++ ++ + ++++ Calcd: 351.11 Found: 352.2  [M + H]⁺ 469

++++ ++ ++ +++ ++ Calcd: 418.18 Found: 419.2  [M + H]⁺ 470

++++ ++ ++ +++ ++ Calcd: 418.18 Found: 419.2  [M + H]⁺ 471

++++ + + ++++ ++ Calcd: 383.12 Found: 384.2  [M + H]⁺ 472

++++ ++ ++ +++ ++ Calcd: 391.16 Found: 392.2  [M + H]⁺ 473

++++ + + ++++ +++ Calcd: 369.10 Found: 370.0  [M + H]⁺ 474

++ + + + Calcd: 379.11 Found: 380.2  [M + H]⁺ 475

++++ + ++ ++++ Calcd: 365.13 Found: 366.2  [M + H]⁺ 476

++++ + + ++++ Calcd: 365.13 Found: 366.2  [M + H]⁺ 477

++++ + ++ +++ Calcd: 413.16 Found: 414.2  [M + H]⁺ 478

++++ ++ +++ +++ +++ Calcd: 413.16 Found: 414.2  [M + H]⁺ 479

++++ + ++ +++ Calcd: 399.14 Found: 400.2  [M + H]⁺ 480

++++ + ++ +++ +++ Calcd: 389.15 Found: 390.2  [M + H]⁺ 481

++++ ++ ++ +++ Calcd: 361.15 Found: 362.0  [M + H]⁺ 482

++++ + ++ +++ Calcd: 361.15 Found: 362.2  [M + H]⁺ 483

++++ ++ + ++ ++ Calcd: 377.15 Found: 378.2 [M + H]⁺ 484

+++ + + + Calcd: 363.13 Found: 364.2  [M + H]⁺ 485

++ + + + Calcd: 379.11 Found: 380.2  [M + H]⁺ 486

++ + + ++ Calcd: 377.15 Found: 378.2  [M + H]⁺ 487

+ + + + Calcd: 362.15 Found: 363.2  [M + H]⁺ 488

+ + + + Calcd: 363.13 Found: 364.2  [M + H]⁺ 489

++ + + + Calcd: 379.11 Found: 380.0  [M + H]⁺ 490

+ + + + Calcd: 362.15 Found: 363.0  [M + H]⁺ 491

+ + + + Calcd: 363.13 Found: 364.2  [M + H]⁺ 492

+++ ++ ++ ++ Calcd: 348.13 Found: 349.2  [M + H]⁺ 493

++++ +++ + ++ Calcd: 399.14 Found: 400.0  [M + H]⁺ 494

++++ ++ + +++ Calcd: 391.13 Found: 392.0  [M + H]⁺ 495

++++ ++ + +++ Calcd: 377.15 Found: 378.2  [M + H]⁺ 496

+++ ++ + ++ Calcd: 363.13 Found: 364.2  [M + H]⁺ 497

+ + + + Calcd: 393.13 Found: 394.0  [M + H]⁺ 498

+++ + + ++ Calcd: 438.16 Found: 439.0  [M + H]⁺ 499

+++ + + ++ Calcd: 405.14 Found: 406.0  [M + H]⁺ 500

+++ + ++ ++ Calcd: 452.18 Found: 452.2  [M + H]⁺ 501

+++ + + ++ Calcd: 397.14 Found: 398.0  [M + H]⁺ 502

++++ + ++ +++ Calcd: 438.16 Found: 439.2  [M + H]⁺ 503

+++ + + ++++ Calcd: 379.14 Found: 380.2  [M + H]⁺ 504

+++ ++ + ++++ Calcd: 389.15 Found: 390.2  [M + H]⁺ 505

+ + + + Calcd: 455.14 Found: 456.0  [M + H]⁺ 506

++++ + + + Calcd: 348.13 Found: 349.2  [M + H]⁺ 507

+ + + + Calcd: 377.15 Found: 378.2  [M + H]⁺ 508

++++ ++ +++ ++ +++ Calcd: 445.22 Found: 446.2  [M + H]⁺ 509

+ + + + Calcd: 439.16 Found: 440.0  [M + H]⁺ 510

++++ ++ +++ ++ ++ Calcd: 431.21 Found: 432.2  [M + H]⁺ 511

++++ ++ +++ ++ ++ Calcd: 431.21 Found: 432.2  [M + H]⁺ 512

++++ ++ +++ ++ +++ Calcd: 445.22 Found: 446.2  [M + H]⁺ 513

+++ + + ++ Calcd: 391.16 Found: 392.2  [M + H]⁺ 514

+++ + + + Calcd: 391.16 Found: 392.2  [M + H]⁺ 515

++++ ++ ++++ +++ +++ Calcd: 445.22 Found: 446.2  [M + H]⁺ 516

+++ + + + Calcd: 382.09 Found: 383.2  [M + H]⁺ 517

+++ + ++ ++ Calcd: 378.14 Found: 379.2 [M + H]⁺ 518

+++ + + + Calcd: 376.16 Found: 377.2  [M + H]⁺ 519

+++ ++ ++ +++ Calcd: 539.18 Found: 540.2  [M + H]⁺ 520

+ + + + Calcd: 441.18 Found: 442.2  [M + H]⁺ 521

++++ ++++ ++++ +++ Calcd: 537.16 Found: 538.0  [M + H]⁺ 522

++++ ++++ ++++ ++++ +++ Calcd: 533.21 Found: 534.2  [M + H]⁺ 523

++++ +++ ++++ ++++ ++ Calcd: 475.14 Found: 476.2  [M + H]⁺ 524

++++ ++++ ++++ ++++ +++ Calcd: 505.18 Found: 506.2  [M + H]⁺ 525

++++ +++ ++++ ++ Calcd: 443.16 Found: 444.2  [M + H]⁺ 526

++++ ++ +++ + ++ Calcd: 447.11 Found: 448.0  [M + H]⁺ 527

++++ ++++ ++++ +++ Calcd: 509.13 Found: 510.2  [M + H]⁺ 528

+++ ++ + + Calcd: 347.14 Found: 348.2  [M + H]⁺ 529

+++ + + ++ Calcd: 371.18 Found: 372.2  [M + H]⁺ *Starting with compound438, proliteration data was obtained using an MDA-MB-361 cell line.

Example 5: Expression and Inhibition Assays of p110α/p85α, p110β/p85α,p110δ/p85α, and p110γ

Commercial kits or systems for assaying PI3-K activities are available.The commercially available kits or systems can be used to screen forinhibitors and/or agonists of PI3-Ks including but not limited to PI3-Kinase α, β, δ, and γ. An exemplary system is PI 3-Kinase (human)HTRF™ Assay from Upstate. The assay can be carried out according to theprocedures suggested by the manufacturer. Briefly, the assay is a timeresolved FRET assay that indirectly measures PIP3 product formed by theactivity of a PI3-K. The kinase reaction is performed in a microtitreplate (e.g., a 384 well microtitre plate). The total reaction volume isapproximately 20 ul per well. In the first step, each well receives 2 ulof test compound in 20% dimethylsulphoxide resulting in a 2% DMSO finalconcentration. Next, approximately 14.5 ul of a kinase/PIP2 mixture(diluted in 1× reaction buffer) is added per well for a finalconcentration of 0.25-0.3 ug/ml kinase and 10 μM PIP2. The plate issealed and incubated for 15 minutes at room temperature. To start thereaction, 3.5 ul of ATP (diluted in 1× reaction buffer) is added perwell for a final concentration of 10 μM ATP. The plate is scaled andincubated for 1 hour at room temperature. The reaction is stopped byadding 5 ul of Stop Solution per well and then 5 ul of Detection Mix isadded per well. The plate is sealed, incubated for 1 hour at roomtemperature, and then read on an appropriate plate reader. Data isanalyzed and IC50s are generated using GraphPad Prism 5. FIG. 1 showsresults for an example analysis of kinase activities. For PI3K α, β, δ,and γ, the nM concentration of inhibitor to reach IC50 is provided.Inhibition of PI3K a at lower concentrations than those for β, δ, and γprovides evidence of specificity within this group of kinases. Similarassays, and others known in the art, can be used to measure the percentinhibition of other kinases, including but not limited to PI3K class IIkinases, phosphoinositide 4 kinases (PI4K), and phosphoinositide 5kinases (PI5K). FIG. 1 provides an analysis of percent inhibition at aconcentration of 1000 nM of the same compound, compound 54, includingPDK class II kinases PI3K2Cβ and PI3KC2γ; the PI4K PI4Kβ; and the PI5sPI5K1α, PI5K1C, PI5k2β, and PI5K2C. Both percent inhibition and IC50 forDYRK1A, HIPK3, and CLK1 kinases are also provided. The results shown inFIG. 1 show the relative degree of specificity of compound 54.

Example 6: Expression and Inhibition Assays of Abl

The cross-activity or lack thereof of one or more compounds of thepresent invention against Abl kinase can be measured according to anyprocedures known in the art or methods disclosed below. For example, thecompounds described herein can be assayed in triplicate againstrecombinant full-length Abl or Abl (T315I) (Upstate) in an assaycontaining 25 mM HEPES, pH 7.4, 10 mM MgCl₂, 200 μM ATP (2.5 μCi ofγ-32P-ATP), and 0.5 mg/mL BSA. The optimized Abl peptide substrateEAIYAAPFAKKK is used as phosphoacceptor (200 μM). Reactions areterminated by spotting onto phosphocellulose sheets, which are washedwith 0.5% phosphoric acid (approximately 6 times, 5-10 minutes each).Sheets are dried and the transferred radioactivity quantitated byphosphorimaging.

Example 7: Expression and Inhibition Assays of Hck

The cross-activity or lack thereof of one or more compounds of thepresent invention against Hck kinase can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be assayed in triplicate against recombinantfull-length Hck in an assay containing 25 mM HEPES, pH 7.4, 10 mM MgCl₂,200 μM ATP (2.5 μCi of γ-32P-ATP), and 0.5 mg/mL BSA. The optimized Srcfamily kinase peptide substrate EIYGEFKKK is used as phosphoacceptor(200 μM). Reactions are terminated by spotting onto phosphocellulosesheets, which are washed with 0.5% phosphoric acid (approximately 6times, 5-10 minutes each). Sheets are dried and the transferredradioactivity quantitated by phosphorimaging.

Example 8: Expression and Inhibition Assays of Inulsin Receptor (IR)

The cross-activity or lack thereof of one or more compounds of thepresent invention against IR receptor kinase can be measured accordingto any procedures known in the art or methods disclosed below. Thecompounds described herein can be assayed in triplicate againstrecombinant insulin receptor kinase domain (Upstate) in an assaycontaining 25 mM HEPES, pH 7.4, 10 mM MgCl₂, 10 mM MnCl₂, 200 μM ATP(2.5 μCi of γ-32P-ATP), and 0.5 mg/mL BSA. Poly E-Y (Sigma; 2 mg/mL) isused as a substrate. Reactions are terminated by spotting ontonitrocellulose, which is washed with 1M NaCl/1% phosphoric acid(approximately 6 times, 5-10 minutes each). Sheets are dried and thetransferred radioactivity quantitated by phosphorimaging.

Example 9: Expression and Inhibition Assays of Src

The cross-activity or lack thereof of one or more compounds of thepresent invention against Src kinase can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be assayed in triplicate against recombinantfull-length Src or Src (T338I) in an assay containing 25 mM HEPES, pH7.4, 10 mM MgCl₂, 200 μM ATP (2.5 μCi of γ-32P-ATP), and 0.5 mg/mL BSA.The optimized Src family kinase peptide substrate EIYGEFKKK is used asphosphoacceptor (200 μM). Reactions are terminated by spotting ontophosphocellulose sheets, which are washed with 0.5% phosphoric acid(approximately 6 times, 5-10 minutes each). Sheets were dried and thetransferred radioactivity quantitated by phosphorimaging.

Example 10: Expression and Inhibition Assays of DNA-PK (DNAK)

The cross-activity or lack thereof of one or more compounds of thepresent invention against DNAK kinase can be measured according to anyprocedures known in the art. DNA-PK can be purchased from Promega andassayed using the DNA-PK Assay System (Promega) according to themanufacturer's instructions.

Example 11: Expression and Inhibition Assays of mTOR

The cross-activity or lack thereof of one or more compounds of thepresent invention against mTor can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be tested against recombinant mTOR (Invitrogen) inan assay containing 50 mM HEPES, pH 7.5, 1 mM EGTA, 10 mM MgCl₂, 2.5 mM,0.01% Tween, 10 μM ATP (2.5 μCi of μ-32P-ATP), and 3 μg/mL BSA. Ratrecombinant PHAS-1/4EBP1 (Calbiochem; 2 mg/mL) is used as a substrate.Reactions are terminated by spotting onto nitrocellulose, which iswashed with 1M NaCl/1% phosphoric acid (approximately 6 times, 5-10minutes each). Sheets are dried and the transferred radioactivityquantitated by phosphorimaging.

Other kits or systems for assaying mTOR activity are commerciallyavailable. For instance, one can use Invitrogen's LanthaScreen™ Kinaseassay to test the inhibitors of mTOR disclosed herein. This assay is atime resolved FRET platform that measures the phosphorylation of GFPlabeled 4EBP1 by mTOR kinase. The kinase reaction is performed in awhite 384 well microtitre plate. The total reaction volume is 20 ul perwell and the reaction buffer composition is 50 mM HEPES pH7.5, 0.01%Polysorbate 20, 1 mM EGTA, 10 mM MnCl₂, and 2 mM DTT. In the first step,each well receives 2 ul of test compound in 20% dimethylsulphoxideresulting in a 2% DMSO final concentration. Next, 8 ul of mTOR dilutedin reaction buffer is added per well for a 60 ng/ml final concentration.To start the reaction, 10 ul of an ATP/GFP-4EBP1 mixture (diluted inreaction buffer) is added per well for a final concentration of 10 μMATP and 0.5 μM GFP-4EBP1. The plate is scaled and incubated for 1 hourat room temperature. The reaction is stopped by adding 10 ul per well ofa Tb-anti-pT46 4EBP1 antibody/EDTA mixture (diluted in TR-FRET buffer)for a final concentration of 1.3 nM antibody and 6.7 mM EDTA. The plateis sealed, incubated for 1 hour at room temperature, and then read on aplate reader set up for LanthaScreen™ TR-FRET. Data is analyzed andIC50s are generated using GraphPad Prism 5.

Example 12: Expression and Inhibition Assays of Vascular EndothelialGrowth Receptor

The cross-activity or lack thereof of one or more compounds of thepresent invention against VEGF receptor can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be tested against recombinant KDR receptor kinasedomain (Invitrogen) in an assay containing 25 mM HEPES. pH 7.4, 10 mMMgCl₂, 0.1% BME, 10 μM ATP (2.5 μCi of μ-32P-ATP), and 3 μg/mL BSA. PolyE-Y (Sigma; 2 mg/mL) is used as a substrate. Reactions are terminated byspotting onto nitrocellulose, which is washed with 1M NaCl/1% phosphoricacid (approximately 6 times, 5-10 minutes each). Sheets are dried andthe transferred radioactivity quantitated by phosphorimaging.

Example 13: Expression and Inhibition Assays of Ephrin Receptor B4(EphB4)

The cross-activity or lack thereof of one or more compounds of thepresent invention against EphB4 can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be tested against recombinant Ephrin receptor B4kinase domain (Invitrogen) in an assay containing 25 mM HEPES, pH 7.4,10 mM MgCl₂, 0.1% BME, 10 μM ATP (2.5 μCi of μ-32P-ATP), and 3 μg/mLBSA. Poly E-Y (Sigma; 2 mg/mL) is used as a substrate. Reactions areterminated by spotting onto nitrocellulose, which is washed with 1MNaCl/1% phosphoric acid (approximately 6 times, 5-10 minutes each).Sheets are dried and the transferred radioactivity quantitated byphosphorimaging.

Example 14: Expression and Inhibition Assays of Epidermal Growth FactorReceptor (EGFR)

The cross-activity or lack thereof of one or more compounds of thepresent invention against EGFR kinase can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be tested against recombinant EGF receptor kinasedomain (Invitrogen) in an assay containing 25 mM HEPES, pH 7.4, 10 mMMgCl₂, 0.1% BME, 10 μM ATP (2.5 μCi of μ-32P-ATP), and 3 μg/mL BSA. PolyE-Y (Sigma; 2 mg/mL) is used as a substrate. Reactions are terminated byspotting onto nitrocellulose, which is washed with 1M NaCl/1% phosphoricacid (approximately 6 times, 5-10 minutes each). Sheets are dried andthe transferred radioactivity quantitated by phosphorimaging.

Example 15: Expression and Inhibition Assays of KIT Assay

The cross-activity or lack thereof of one or more compounds of thepresent invention against KIT kinase can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be tested against recombinant KIT kinase domain(Invitrogen) in an assay containing 25 mM HEPES, pH 7.4, 10 mM MgCl₂, 1mM DTT, 10 mM MnCl₂, 10 μM ATP (2.5 μCi of μ-32P-ATP), and 3 μg/mL BSA.Poly E-Y (Sigma; 2 mg/mL) is used as a substrate. Reactions areterminated by spotting onto nitrocellulose, which is washed with 1MNaCl/1% phosphoric acid (approximately 6 times, 5-10 minutes each).Sheets are dried and the transferred radioactivity quantitated byphosphorimaging.

Example 16: Expression and Inhibition Assays of RET

The cross-activity or lack thereof of one or more compounds of thepresent invention against RET kinase can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be tested against recombinant RET kinase domain(Invitrogen) in an assay containing 25 mM HEPES, pH 7.4, 10 mM MgCl₂,2.5 mM DTT, 10 μM ATP (2.5 μCi of μ-32P-ATP), and 3 μg/mL BSA. Theoptimized Abl peptide substrate EAIYAAPFAKKK is used as phosphoacceptor(200 μM). Reactions are terminated by spotting onto phosphocellulosesheets, which are washed with 0.5% phosphoric acid (approximately 6times, 5-10 minutes each). Sheets are dried and the transferredradioactivity quantitated by phosphorimaging.

Example 17: Expression and Inhibition Assays of Platelet Derived GrowthFactor Receptor (PDGFR)

The cross-activity or lack thereof of one or more compounds of thepresent invention against PDGFR kinase can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be tested against recombinant PDG receptor kinasedomain (Invitrogen) in an assay containing 25 mM HEPES, pH 7.4, 10 mMMgCl₂, 2.5 mM DTT, 10 μM ATP (2.5 μCi of μ-32P-ATP), and 3 μg/mL BSA.The optimized Abl peptide substrate EAIYAAPFAKKK is used asphosphoacceptor (200 μM). Reactions are terminated by spotting ontophosphocellulose sheets, which are washed with 0.5% phosphoric acid(approximately 6 times, 5-10 minutes each). Sheets are dried and thetransferred radioactivity quantitated by phosphorimaging.

Example 18: Expression and Inhibition Assays of FMS-Related TyrosineKinase 3 (FLT-3)

The cross-activity or lack thereof of one or more compounds of thepresent invention against FLT-3 kinase can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be tested against recombinant FLT-3 kinase domain(Invitrogen) in an assay containing 25 mM HEPES, pH 7.4, 10 mM MgCl₂,2.5 mM DTT, 10 μM ATP (2.5 μCi of μ-32P-ATP), and 3 μg/mL BSA. Theoptimized Abl peptide substrate EAIYAAPFAKKK is used as phosphoacceptor(200 μM). Reactions are terminated by spotting onto phosphocellulosesheets, which are washed with 0.5% phosphoric acid (approximately 6times, 5-10 minutes each). Sheets are dried and the transferredradioactivity quantitated by phosphorimaging.

Example 19: Expression and Inhibition Assays of TEK Receptor TyrosineKinase (TIE2)

The cross-activity or lack thereof of one or more compounds of thepresent invention against TIE2 kinase can be measured according to anyprocedures known in the art or methods disclosed below. The compoundsdescribed herein can be tested against recombinant TIE2 kinase domain(Invitrogen) in an assay containing 25 mM HEPES, pH 7.4, 10 mM MgCl₂, 2mM DTT, 10 mM MnCl₂, 10 μM ATP (2.5 μCi of μ-32P-ATP), and 3 μg/mL BSA.Poly E-Y (Sigma; 2 mg/mL) is used as a substrate. Reactions areterminated by spotting onto nitrocellulose, which is washed with 1MNaCl/1% phosphoric acid (approximately 6 times, 5-10 minutes each).Sheets are dried and the transferred radioactivity quantitated byphosphorimaging.

Example 20: B Cell Activation and Proliferation Assay

The ability of one or more subject compounds to inhibit B cellactivation and proliferation is determined according to standardprocedures known in the art. For example, an in vitro cellularproliferation assay is established that measures the metabolic activityof live cells. The assay is performed in a 96 well microtiter plateusing Alamar Blue reduction. Balb/c splenic B cells are purified over aFicoll-Paque™ PLUS gradient followed by magnetic cell separation using aMACS B cell Isolation Kit (Miletenyi). Cells are plated in 90 ul at50,000 cells/well in B Cell Media (RPMI+10% FBS+Penn/Strep+50 μM bME+5mM HEPES). A compound disclosed herein is diluted in B Cell Media andadded in a 10 ul volume. Plates are incubated for 30 min at 37 C and 5%CO₂ (0.2% DMSO final concentration). A 50 ul B cell stimulation cocktailis then added containing either 10 ug/ml LPS or 5 ug/ml F(ab′)2 Donkeyanti-mouse IμM plus 2 ng/ml recombinant mouse IL4 in B Cell Media.Plates are incubated for 72 hours at 37° C. and 5% CO₂. A volume of 15μL of Alamar Blue reagent is added to each well and plates are incubatedfor 5 hours at 37 C and 5% CO₂. Alamar Blue fluoresce is read at560Ex/590Em, and IC50 or EC50 values are calculated using CiraphPadPrism 5.

Example 21: Tumor Cell Line Proliferation Assay

The ability of one or more subject compounds to inhibit tumor cell lineproliferation is determined according to standard procedures known inthe art. For instance, an in vitro cellular proliferation assay can beperformed to measure the metabolic activity of live cells. The assay isperformed in a 96 well microtiter plate using Alamar Blue reduction.Human tumor cell lines are obtained from ATCC (e.g., MCF7, U-87 MG,MDA-MB-468, PC-3), grown to confluency in T75 flasks, trypsinized with0.25% trypsin, washed one time with Tumor Cell Media (DMEM+10% FBS), andplated in 90 ul at 5,000 cells/well in Tumor Cell Media. A compounddisclosed herein is diluted in Tumor Cell Media and added in a 10 ulvolume. Plates are incubated for 72 hours at 37 C and 5% CO₂. A volumeof 10 uL of Alamar Blue reagent is added to each well and plates areincubated for 3 hours at 37 C and 5% CO₂. Alamar Blue fluoresce is readat 560Ex/590Em, and IC50 values are calculated using GraphPad Prism 5.The results are expected to show that some of the compounds of thepresent invention are potent inhibitors of tumor cell line proliferationunder the conditions tested.

Example 22: Antitumor Activity In Vivo

The compounds described herein can be evaluated in a panel of human andmurine tumor models.

Paclitaxel-Refractory Tumor Models

1. Clinically-derived Ovarian Carcinoma Model.

This tumor model is established from a tumor biopsy of an ovarian cancerpatient. Tumor biopsy is taken from the patient.

The compounds described herein are administered to nude mice bearingstaged tumors using an every 2 days×5 schedule.

2. A2780Tax Human Ovarian Carcinoma Xenograft (Mutated Tubulin).

A2780Tax is a paclitaxel-resistant human ovarian carcinoma model. It isderived from the sensitive parent A2780 line by co-incubation of cellswith paclitaxel and verapamil, an MDR-reversal agent. Its resistancemechanism has been shown to be non-MDR related and is attributed to amutation in the gene encoding the beta-tubulin protein.

The compounds described herein can be administered to mice bearingstaged tumors on an every 2 days×5 schedule.

3. HCT116NM46 Human Colon Carcinoma Xenograft (Multi-Drug Resistant).

HCT116NM46 is an MDR-resistant colon carcinoma developed from thesensitive HCT116 parent line. In vivo, grown in nude mice, HCT116/VM46has consistently demonstrated high resistance to paclitaxel.

The compounds described herein can be administered to mice bearingstaged tumors on an every 2 days×5 schedule.

5. M5076 Murine Sarcoma Model

M5076 is a mouse fibrosarcoma that is inherently refractory topaclitaxel in vivo.

The compounds described herein can be administered to mice bearingstaged tumors on an every 2 days×5 schedule.

One or more compounds of the invention can be used in combination othertherapeutic agents in vivo in the multidrug resistant human coloncarcinoma xenografts HCT/VM46 or any other model known in the artincluding those described herein.

The results are expected to show that one or more compounds of thepresent invention are potent inhibitors of tumor growth in vivo underthe conditions tested.

Example 23: Microsome Stability Assay

The stability of one or more subject compounds is determined accordingto standard procedures known in the art. For example, stability of oneor more subject compounds is established by an in vitro assay. Inparticular, an in vitro microsome stability assay is established thatmeasures stability of one or more subject compounds when reacting withmouse, rat or human microsomes from liver. The microsome reaction withcompounds is performed in 1.5 mL Eppendorf tube. Each tube contains 0.1μL of 10.0 mg/ml NADPH; 75 μL of 20.0 mg/ml mouse, rat or human livermicrosome; 0.4 μL of 0.2 M phosphate buffer, and 425 μL of ddH₂O.Negative control (without NADPH) tube contains 75 μL of 20.0 mg/mlmouse, rat or human liver microsome; 0.4 μL of 0.2 M phosphate buffer,and 525 μL of ddH₂O. The reaction is started by adding 1.0 μL of 10.0 mMtested compound. The reaction tubes are incubated at 37° C. 100 μLsample is collected into new Eppendorf tube containing 300 μL coldMethanol at 0, 5, 10, 15, 30 and 60 minutes of reaction. Samples arecentrifuged at 15,000 rpm to remove protein. Supernatant of centrifugedsample is transferred to new tube. Concentration of stable compoundafter reaction with microsome in the supernatant is measured by LiquidChromatography/Mass Spectrometry (LC-MS).

Example 24: Plasma Stability Assay

The stability of one or more subject compounds in plasma is determinedaccording to standard procedures known in the art. See, e.g., RapidCommun. Mass Spectrom., 10: 1019-1026. The following procedure is anHPLC-MS/MS assay using human plasma; other species including monkey,dog, rat, and mouse are also available. Frozen, heparinized human plasmais thawed in a cold water bath and spun for 10 minutes at 2000 rpm at 4°C. prior to use. A subject compound is added from a 400 μM stocksolution to an aliquot of pre-warmed plasma to give a final assay volumeof 400 μL (or 800 μL for half-life determination), containing 5 μM testcompound and 0.5% DMSO. Reactions are incubated, with shaking, for 0minutes and 60 minutes at 37° C., or for 0, 15, 30, 45 and 60 minutes at37 C for half life determination. Reactions are stopped by transferring50 μL of the incubation mixture to 200 μL of ice-cold acetonitrile andmixed by shaking for 5 minutes. The samples are centrifuged at 6000×gfor 15 minutes at 4° C. and 120 μL of supernatant removed into cleantubes. The samples are then evaporated to dryness and submitted foranalysis by HPLC-MS/MS.

Where desired, one or more control or reference compounds (5 μM) aretested simultaneously with the test compounds: one compound,propoxycaine, with low plasma stability and another compound,propantheline, with intermediate plasma stability.

Samples are reconstituted in acetonitrile/methanol/water (1/1/2, v/v/v)and analyzed via (RP)HPLC-MS/MS using selected reaction monitoring(SRM). The HPLC conditions consist of a binary LC pump with autosampler,a mixed-mode, C12, 2×20 mm column, and a gradient program. Peak areascorresponding to the analytes are recorded by HPLC-MS/MS. The ratio ofthe parent compound remaining after 60 minutes relative to the amountremaining at time zero, expressed as percent, is reported as plasmastability. In case of half-life determination, the half-life isestimated from the slope of the initial linear range of the logarithmiccurve of compound remaining (%) vs. time, assuming first order kinetics.

Example 25: Chemical Stability

The chemical stability of one or more subject compounds is determinedaccording to standard procedures known in the art. The following detailsan exemplary procedure for ascertaining chemical stability of a subjectcompound. The default buffer used for the chemical stability assay isphosphate-buffered saline (PBS) at pH 7.4; other suitable buffers can beused. A subject compound is added from a 100 μM stock solution to analiquot of PBS (in duplicate) to give a final assay volume of 400 μL,containing 5 μM test compound and 1% DMSO (for half-life determination atotal sample volume of 700 μL is prepared). Reactions are incubated,with shaking, for 0 minutes and 24 hours at 37° C.; for half-lifedetermination samples are incubated for 0, 2, 4, 6, and 24 hours.Reactions are stopped by adding immediately 100 μL of the incubationmixture to 100 μL of acetonitrile and vortexing for 5 minutes. Thesamples are then stored at −20° C. until analysis by HPLC-MS/MS. Wheredesired, a control compound or a reference compound such as chlorambucil(5 μM) is tested simultaneously with a subject compound of interest, asthis compound is largely hydrolyzed over the course of 24 hours. Samplesare analyzed via (RP)HPLC-MS/MS using selected reaction monitoring(SRM). The HPLC conditions consist of a binary LC pump with autosampler,a mixed-mode, C12, 2×20 mm column, and a gradient program. Peak areascorresponding to the analytes are recorded by HPLC-MS/MS. The ratio ofthe parent compound remaining after 24 hours relative to the amountremaining at time zero, expressed as percent, is reported as chemicalstability. In case of half-life determination, the half-life isestimated from the slope of the initial linear range of the logarithmiccurve of compound remaining (%) vs. time, assuming first order kinetics.

Example 26: Akt Kinase Assay

Cells comprising components of the Akt/mTOR pathway, including but notlimited to L6 myoblasts, B-ALL cells, B-cells, T-cells, leukemia cells,bone marrow cells, p190 transduced cells, philladelphia chromosomepositive cells (Ph+), and mouse embryonic fibroblasts, are typicallygrown in cell growth media such as DMEM supplemented with fetal bovinescrum and/or antibiotics, and grown to confluency.

In order to compare the effect of one or more compounds disclosed hereinon Akt activation, said cells are serum starved overnight and incubatedwith one or more compounds disclosed herein or about 0.1% DMSO forapproximately 1 minute to about 1 hour prior to stimulation with insulin(e.g., 100 nM) for about 1 minutes to about 1 hour. Cells are lysed byscraping into ice cold lysis buffer containing detergents such as sodiumdodecyl sulfate and protease inhibitors (e.g., PMSF). After contactingcells with lysis buffer, the solution is briefly sonicated, cleared bycentrifugation, resolved by SDS-PAGE, transferred to nitrocellulose orPVDF and immunoblotted using antibodies to phospho-Akt S473, phospho-AktT308, Akt, and 3-actin (Cell Signaling Technologies).

Example 27: Kinase Signaling in Blood

PI3K/Akt/mTor signaling is measured in blood cells using the phosflowmethod (Methods Enzymol. 2007; 434:131-54). The advantage of this methodis that it is by nature a single cell assay so that cellularheterogeneity can be detected rather than population averages. Thisallows concurrent distinction of signaling states in differentpopulations defined by other markers. Phosflow is also highlyquantitative. To test the effects of one or more compounds disclosedherein, unfractionated splenocytes, or peripheral blood mononuclearcells are stimulated with anti-CD3 to initiate T-cell receptorsignaling. The cells are then fixed and stained for surface markers andintracellular phosphoproteins. It is expected that inhibitors disclosedherein inhibit anti-CD3 mediated phosphorylation of Akt-S473 and S6,whereas rapamycin inhibits S6 phosphorylation and enhances Aktphosphorylation under the conditions tested.

Similarly, aliquots of whole blood are incubated for 15 minutes withvehicle (e.g., 0.1% DMSO) or kinase inhibitors at variousconcentrations, before addition of stimuli to crosslink the T cellreceptor (TCR) (anti-CD3 with secondary antibody) or the B cell receptor(BCR) using anti-kappa light chain antibody (Fab′2 fragments). Afterapproximately 5 and 15 minutes, samples are fixed (e.g., with cold 4%paraformaldehyde) and used for phosflow. Surface staining is used todistinguish T and B cells using antibodies directed to cell surfacemarkers that are known to the art. The level of phosphrylation of kinasesubstrates such as Akt and S6 are then measured by incubating the fixedcells with labeled antibodies specific to the phosphorylated isoforms ofthese proteins. The population of cells is then analyzed by flowcytometry.

The results are expected to show that one or more of the compounds ofthe present invention are potent and selective inhibitors of one or moremembers of one or more of PI3K, mTOR, and Akt signaling in blood cellsunder the conditions tested.

Example 28: Colony Formation Assay

Murine bone marrow cells freshly transformed with a p190 BCR-Ablretrovirus (herein referred to as p190 transduced cells) are plated inthe presence of various drug combinations in M3630 methylcellulose mediafor about 7 days with recombinant human IL-7 in about 30% serum, and thenumber of colonies formed is counted by visual examination under amicroscope.

Alternatively, human peripheral blood mononuclear cells are obtainedfrom Philadelphia chromosome positive (Ph+) and negative (Ph−) patientsupon initial diagnosis or relapse. Live cells are isolated and enrichedfor CD19+CD34+ B cell progenitors. After overnight liquid culture, cellsare plated in methocult GF+ H4435, Stem Cell Technologies) supplementedwith cytokines (IL-3, IL-6, IL-7, G-CSF, GM-CSF, CF, Flt3 ligand, anderythropoietin) and various concentrations of known chemotherapeuticagents in combination with either compounds of the present disclosure.Colonies are counted by microscopy 12-14 days later. This method can beused to test for evidence of additive or synergistic activity.

The results are expected to show that one or more the compounds of thepresent invention are potent and selective inhibitors of p190 transducedcell colony formation under the conditions tested.

Example 29: In Vivo Effect of Kinase Inhibitors on Leukemic Cells

Female recipient mice are lethally irradiated from a 7 source in twodoses about 4 hr apart, with approximately 5Gy each. About 1 hr afterthe second radiation dose, mice are injected i.v. with about 1×10⁶leukemic cells (e.g., Ph+ human or murine cells, or p190 transduced bonemarrow cells). These cells are administered together with aradioprotective dose of about 5×10⁶ normal bone marrow cells from 3-5week old donor mice. Recipients are given antibiotics in the water andmonitored daily. Mice who become sick after about 14 days are euthanizedand lymphoid organs are harvested for analysis. Kinase inhibitortreatment begins about ten days after leukemic cell injection andcontinues daily until the mice become sick or a maximum of approximately35 days post-transplant. Inhibitors are given by oral lavage.

Peripheral blood cells are collected approximately on day 10(pre-treatment) and upon euthanization (post treatment), contacted withlabeled anti-hCD4 antibodies and counted by now cytometry. This methodcan be used to demonstrate that the synergistic effect of one or morecompounds disclosed herein in combination with known chemotherapeuticagents significantly reduce leukemic blood cell counts as compared totreatment with known chemotherapeutic agents (e.g., Gleevec) alone underthe conditions tested.

Example 30: Treatment of Lupus Disease Model Mice

Mice lacking the inhibitory receptor FcγRIIb that opposes PI3K signalingin B cells develop lupus with high penetrance. FcγRIIb knockout mice(R2KO, Jackson Labs) are considered a valid model of the human diseaseas some lupus patients show decreased expression or function of FcγRIIb(S. Bolland and J. V. Ravtech 2000. Immunity 12:277-285).

The R2KO mice develop lupus-like disease with anti-nuclear antibodies,glomerulonephritis and proteinurea within about 4-6 months of age. Forthese experiments, the rapamycin analogue RAD001 (available from LCLaboratories) is used as a benchmark compound, and administered orally.This compound has been shown to ameliorate lupus symptoms in theB6.Sle1z.Sle3z model (T. Wu et al. J. Clin Invest. 117:2186-2196).

Lupus disease model mice such as R2KO, BXSB or MLR/lpr are treated atabout 2 months old, approximately for about two months. Mice are givendoses of: vehicle, RAD001 at about 10 mg/kg, or compounds disclosedherein at approximately 1 mg/kg to about 500 mg/kg. Blood and urinesamples are obtained at approximately throughout the testing period, andtested for antinuclear antibodies (in dilutions of serum) or proteinconcentration (in urine). Serum is also tested for anti-ssDNA andanti-dsDNA antibodies by ELISA. Animals are euthanized at day 60 andtissues harvested for measuring spleen weight and kidney disease.Glomerulonephritis is assessed in kidney sections stained with H&E.Other animals are studied for about two months after cessation oftreatment, using the same endpoints.

This model established in the art can be employed to test that thekinase inhibitors disclosed herein can suppress or delay the onset oflupus symptoms in lupus disease model mice.

Example 31: Murine Bone Marrow Transplant Assay

Female recipient mice are lethally irradiated from a γ ray source. About1 hr after the radiation dose, mice are injected with about 1×106leukemic cells from carly passage p190 transduced cultures (e.g., asdescribed in Cancer Genet Cytogenet. 2005 August; 161(1):51-6). Thesecells are administered together with a radioprotective dose ofapproximately 5×106 normal bone marrow cells from 3-5 wk old donor mice.Recipients are given antibiotics in the water and monitored daily. Micewho become sick after about 14 days are euthanized and lymphoid organsharvested for flow cytometry and/or magnetic enrichment. Treatmentbegins on approximately day 10 and continues daily until mice becomesick, or after a maximum of about 35 days post-transplant. Drugs aregiven by oral gavage (p.o.). In a pilot experiment a dose ofchemotherapeutic that is not curative but delays leukemia onset by aboutone week or less is identified; controls are vehicle-treated or treatedwith chemotherapeutic agent, previously shown to delay but not cureleukemogenesis in this model (e.g., imatinib at about 70 mg/kg twicedaily). For the first phase p190 cells that express eGFP are used, andpostmortem analysis is limited to enumeration of the percentage ofleukemic cells in bone marrow, spleen and lymph node (LN) by flowcytometry. In the second phase, p190 cells that express a tailless formof human CD4 are used and the postmortem analysis includes magneticsorting of hCD4+ cells from spleen followed by immunoblot analysis ofkey signaling endpoints: p Akt-T308 and S473; pS6 and p4EBP-1. Ascontrols for immunoblot detection, sorted cells are incubated in thepresence or absence of kinase inhibitors of the present disclosureinhibitors before lysis. Optionally, “phosflow” is used to detect pAkt-S473 and pS6-S235/236 in hCD4-gated cells without prior sorting.These signaling studies are particularly useful if, for example,drug-treated mice have not developed clinical leukemia at the 35 daytime point. Kaplan-Meier plots of survival are generated and statisticalanalysis done according to methods known in the art. Results from p190cells are analyzed separated as well as cumulatively.

Samples of peripheral blood (100-200 μl) are obtained weekly from allmice, starting on day 10 immediately prior to commencing treatment.Plasma is used for measuring drug concentrations, and cells are analyzedfor leukemia markers (eGFP or hCD4) and signaling biomarkers asdescribed herein.

This general assay known in the art may be used to test that effectivetherapeutic doses of the compounds disclosed herein can be used forinhibiting the proliferation of leukemic cells.

Example 32: Rat Developing Type II Collagen Induced Arthritis Assay

In order to study the effects of the compounds of the present inventionon the autoimmune disease arthritis, a collagen induced developingarthritis model is used. Female Lewis rats are given collagen injectionsat day 0. Bovine type II collagen is prepared as a 4 mg/ml solution in0.01N acetic acid. Equal volumes of collagen and Freund's incompleteadjuvant are emulsified by hand mixing until a bead of the emulsifiedmaterial holds its form in water. Each rodent receives a 300 μlinjection of the mixture at each injection time spread over threesubcutaneous sites on the back.

Oral compound administration begins on day 0 and continues through day16 with vehicle (5% NMP, 85% PEG 400, 10% Solutol) or compounds of thepresent invention in vehicle or control (e.g., methotrexate) at 12 hourintervals daily. Rats are weighed on days 0, 3, 6, 9-17 and calipermeasurements of ankles are taken on days 9-17. Final body weights aretaken, and then the animals are euthanized on day 17. Aftereuthanization, blood is drawn and hind paws and knees are removed. Bloodis further processed for pharmacokinetics experiments as well as ananti-type II collagen antibody ELISA assay. Hind paws are weighed andthen, with the knees, preserved in 10% formalin. The paws and knees aresubsequently processed for microscopy. Livers, spleen and thymus areweighed. Sciatic nerves are prepared for histopathology.

Knee and ankle joints are fixed for 1-2 days and decalcified for 4-5days. Ankle joints are cut in half longitudinally, and knees are cut inhalf along the frontal plane. Joints are processed, embedded, sectionedand stained with toluidine blue. Scoring of the joints is done accordingto the following criteria:

Knee and Ankle Inflammation 0=Normal

1=Minimal infiltration of inflammatory cells in synovium/periarticulartissue2=Mild infiltration3=Moderate infiltration with moderate edema4=Marked infiltration with marked edema5=Severe infiltration with severe edema

Ankle Pannus 0=Normal

1=Minimal infiltration of pannus in cartilage and subchondral bone2=Mild infiltration (<¼ of tibia or tarsals at marginal zones)3=Moderate infiltration (¼ to ⅓ of tibia or small tarsals affected atmarginal zones)4=Marked infiltration (½=¾ of tibia or tarsals affected at marginalzones)5=Severe infiltration (>¾ of tibia or tarsals affected at marginalzones, severe distortion of overall architecture)

Knee Pannus 0=Normal

1=Minimal infiltration of pannus in cartilage and subchondral bone2=Mild infiltration (extends over up to ¼ of surface or subchondral areaof tibia or femur)3=Moderate infiltration (extends over >¼ but <½ of surface orsubchondral area of tibia or femur)4=Marked infiltration (extends over ½ to ¾ of tibial or femoral surface)5=Severe infiltration (covers >¾ of surface)

Cartilage Damage (Ankle, Emphasis on Small Tarsals) 0=Normal

1=Minimal=minimal to mild loss of toluidine blue staining with noobvious chondrocyte loss or collagen disruption2=Mild=mild loss of toluidine blue staining with focal mild(superficial) chondrocyte loss and/or collagen disruption3=Moderate=moderate loss of toluidine blue staining with multifocalmoderate (depth to middle zone) chondrocyte loss and/or collagendisruption, smaller tarsals affected to ½=¾ depth4=Marked=marked loss of toluidine blue staining with multifocal marked(depth to deep zone) chondrocyte loss and/or collagen disruption, 1 ormore small tarsals have full thickness loss of cartilage5=Severe=severe diffuse loss of toluidine blue staining with multifocalsevere (depth to tide mark) chondrocyte loss and/or collagen disruption

Cartilage Damage (Knee, Emphasis on Femoral Condyles) 0=Normal

1=Minimal=minimal to mild loss of toluidine blue staining with noobvious chondrocyte loss or collagen disruption2=Mild=mild loss of toluidine blue staining with focal mild(superficial) chondrocyte loss and/or collagen disruption3=Moderate=moderate loss of toluidine blue staining with multifocal todiffuse moderate (depth to middle zone) chondrocyte loss and/or collagendisruption4=Marked=marked loss of toluidine blue staining with multifocal todiffuse marked (depth to deep zone) chondrocyte loss and/or collagendisruption or single femoral surface with total or near total loss5=Severe=severe diffuse loss of toluidine blue staining with multifocalsevere (depth to tide mark) chondrocyte loss and/or collagen disruptionon both femurs and/or tibias

Bone Resorption (Ankle) 0=Normal

1=Minimal=small areas of resorption, not readily apparent on lowmagnification, rare osteoclasts2=Mild=more numerous areas of resorption, not readily apparent on lowmagnification, osteoclasts more numerous, <¼ of tibia or tarsals atmarginal zones resorbed3=Moderate=obvious resorption of medullary trabecular and cortical bonewithout full thickness defects in cortex, loss of some medullarytrabeculae, lesion apparent on low magnification, osteoclasts morenumerous, ¼ to ⅓ of tibia or tarsals affected at marginal zones4=Marked=Full thickness defects in cortical bone, often with distortionof profile of remaining cortical surface, marked loss of medullary bone,numerous osteoclasts, ½=¾ of tibia or tarsals affected at marginal zones5=Severe=Full thickness defects in cortical bone, often with distortionof profile of remaining cortical surface, marked loss of medullary bone,numerous ostcoclasts, >¾ of tibia or tarsals affected at marginal zones,severe distortion of overall architecture

Bone Resorption (Knee) 0=Normal

1=Minimal=small areas of resorption, not readily apparent on lowmagnification, rare ostcoclasts2=Mild=more numerous areas of resorption, definite loss of subchondralbone involving ¼ of tibial or femoral surface (medial or lateral)3=Moderate=obvious resorption of subchondral bone involving >¼ but <½ oftibial or femoral surface (medial or lateral)4=Marked=obvious resorption of subchondral bone involving >½ but <¾ oftibial or femoral surface (medial or lateral)5=Severe=distortion of entire joint due to destruction involving >¾ oftibial or femoral surface (medial or lateral)

Statistical analysis of body/paw weights, paw AUC parameters andhistopathologic parameters were evaluated using a Student's t-test orother appropriate (ANOVA with post-test) with significance set at the 5%significance level. Percent inhibition of paw weight and AUC wascalculated using the following formula:

% Inhibition=A−B/Aλ100

A=Mean Disease Control−Mean Normal

B=Mean Treated−Mean Normal

The results are expected to show, relative to vehicle only control or tomethotrexate control, that the compounds of the present inventionexhibit a significant reduction in arthritis induced ankle diameterincrease over time, and reduction of ankle histopathology in at leastone or more of the categories of inflammation, pannus, cartilage damage,and bone resorption as described above. The results are expected to showthat one or more compounds of the present invention may be useful forthe treatment and reduction of arthritis disease symptoms.

The results further are expected to show a reduction at 10, 20, and 60mg/kg dosage levels of serum anti-type II collagen levels for selectedtest compounds, suggesting that one or more compounds of the presentinvention may not only be useful for the treatment and reduction ofarthritis disease symptoms, but may also be useful for the inhibition ofthe autoimmune reaction itself.

Example 33: Rat Established Type II Collagen Induced Arthritis Assay

In order to examine the dose responsive efficacy of the compounds of thepresent invention in inhibiting the inflammation, cartilage destructionand bone resorption of 10 day established type II collagen inducedarthritis in rats, compounds are administered orally daily or twicedaily for 6 days.

Female Lewis rats are anesthetized and given collagen injectionsprepared and administered as described previously on day 0. On day 6,animals are anesthetized and given a second collagen injection. Calipermeasurements of normal (pre-disease) right and left ankle joints areperformed on day 9. On days 10-11, arthritis typically occurs and ratsare randomized into treatment groups. Randomization is performed afterankle joint swelling is obviously established and there is evidence ofbilateral disease.

After an animal is selected for enrollment in the study, treatment isinitiated by the oral route. Animals are given vehicle, control (Enbrel)or compound doses, twice daily or once daily (BID or QD respectively).Administration is performed on days 1-6 using a volume of 2.5 ml/kg(BID) or 5 ml/kg (QD) for oral solutions. Rats are weighed on days 1-7following establishment of arthritis and caliper measurements of anklestaken every day. Final body weights are taken on day 7 and animals areeuthanized.

The results are expected to show reduction in mean ankle diameterincrease over time for selected test compounds under the conditionstested.

Example 34: Adjuvant Induced Arthritis Assay Intrathecal Catheterizationof Rats

Isoflurane-anesthetized Lewis rats (200-250 g) are implanted with anintrathecal (IT) catheter. After a 6 d recovery period, all animalsexcept those that appeared to have sensory or motor abnormalities(generally fewer than 5% of the total number) are used for experiments.For IT administration, 10 μl of drug or saline followed by 10 μl ofisotonic saline is injected through the catheter.

Adjuvant Arthritis and Drug Treatment

Lewis rats are immunized at the base of the tail with 0.1 ml of completeFreund's adjuvant (CFA) on day 0 several days after catheterimplantation (n=6/group). Drug (e.g., one or more compounds of thepresent invention or vehicle) treatment is generally started on day 8and is continued daily until day 20. Clinical signs of arthritisgenerally begin on day 10, and paw swelling is determined every secondday by water displacement plethysmometry.

The results are expected to show that one or more compounds of thepresent invention demonstrates may be useful for the treatment of one ormore of the diseases or conditions described herein.

Example 35: Rodent Pharmacokinetic Assay

In order to study the pharmacokinetics of the compounds of the presentinvention a set of 4-10 week old mice are grouped according to thefollowing table:

Compound Administration Group# Mice/group (mg/kg) Route Regimen 1 3 1 PoOne dose 2 3 3 3 3 10 4 3 30 5 3 60

Compounds of the present invention are dissolved in an appropriatevehicle (e.g., 5% 1-methyl-2-pyrrolidinone, 85% polyethylene glycol 400,10% Solutor) and administered orally at 12 hour intervals daily. Allanimals are euthanized in CO₂ 2 hours after the final compound isadministered. Blood is collected immediately and kept on ice for plasmaisolation. Plasma is isolated by centrifuging at 5000 rpm for 10minutes. Harvested plasma is frozen for pharmacokinetic detection.

The results are expected to demonstrate the pharmacokinetic parameterssuch as absorption, distribution, metabolism, excretion, and toxicityfor the compounds of the present invention.

Example 36: Basotest Assay

The basotest assay is performed using Orpegen Pharma Basotest reagentkit. Heparinized whole blood is pre-incubated with test compound orsolvent at 37 C for 20 min. Blood is then incubated with assay kitstimulation buffer (to prime cells for response) followed by allergen(dust mite extract or grass extract) for 20 min. The degranulationprocess is stopped by incubating the blood samples on ice. The cells arethen labeled with anti-IgE-PE to detect basophilic granulocytes, andanti-gp53-FITC to detect gp53 (a glycoprotein expressed on activatedbasophils). After staining red blood cells are lysed by addition ofLysing Solution. Cells are washed, and analyzed by flow cytometry. Testcompounds, when evaluated in this assay inhibit allergen inducedactivation of basophilic granulocytes at sub micromolar range. Theresults are expected to demonstrate that under the conditions tested oneor more compounds of the present invention are capable of inhibitingallergen induced activation of basophils.

Example 37: Use of the Compounds of the Present Invention for Inhibitionof Tumor Growth Cell Lines

Cell lines of interest (A549, U87, ZR-75-1 and 786-O) are obtained fromAmerican Type Culture Collection (ATCC, Manassas, Va.). Cells areproliferated and preserved cryogenically at early passage (e.g., passage3). One aliquot is used for further proliferation to get enough cellsfor one TGI study (at about passage 9).

Animals

Female athymic nude mice are supplied by Harlan. Mice are received at 4to 6 weeks of age. All mice are acclimated for about one day to twoweeks prior to handling. The mice are housed in microisolator cages andmaintained under specific pathogen-free conditions. The mice are fedwith irradiated mouse chow and freely available autoclaved water isprovided.

Tumor Xenograft Model

Mice are inoculated subcutaneously in the right flank with 0.01 to 0.5ml of tumor cells (approximately 1.0×10⁵ to 1.0×10⁸ cells/mouse). Fiveto 10 days following inoculation, tumors are measured using calipers andtumor weight is calculated, for example using the animal studymanagement software, such as Study Director V.1.6.70 (Study Log). Micewith tumor sizes of about 120 mg are pair-matched into desired groupsusing Study Director (Day 1). Body weights are recorded when the miceare pair-matched. Tumor volume and bodyweight measurements are taken oneto four times weekly and gross observations are made at least oncedaily. On Day 1, compounds of the present invention and referencecompounds as well as vehicle control are administered by oral gavage oriv as indicated. At the last day of the experiment, mice are sacrificedand their tumors are collected 1-4 hours after the final dose. Thetumors are excised and cut into two sections. One third of the tumor isfixed in formalin and embedded in paraffin blocks and the remaining twothirds of tumor is snap frozen and stored at −80° C.

Data and Statistical Analysis

Mean tumor growth inhibition (TGI) is calculated utilizing the followingformula:

${T\; G\; I} = {\lbrack {1 - \frac{( {{\overset{\_}{\chi}}_{{Treated}_{({Final})}} - {\overset{\_}{\chi}}_{{Treated}_{({{Day}\; 1})}}} )}{( {{\overset{\_}{\chi}}_{{Control}_{({Final})}} - {\overset{\_}{\chi}}_{{Control}_{({{Day}\; 1})}}} }} \rbrack \times 100\%}$

Tumors that regress from the Day 1 starting size are removed from thecalculations. Individual tumor shrinkage (TS) is calculated using theformula below for tumors that show regression relative to Day 1 tumorweight. The mean tumor shrinkage of each group is calculated andreported.

${T\; S} = {\lbrack {1 - \frac{( {{Tumor}\mspace{14mu} {Weight}_{({Final})}} )}{( {{Tumor}\mspace{14mu} {Weight}_{({{Day}\; 1})}} )}} \rbrack \times 100\%}$

The model can be employed to show whether the compounds of the presentinvention can inhibit tumor cell growth such as renal carcinoma cellgrowth, breast cancer cell growth, lung cancer cell growth, orglioblastoma cell growth under the conditions tested.

Example 38: Inhibition of PI3K Pathway and Proliferation of Tumor Cellswith PI3Kα Mutation

Cells comprising one or more mutations in PI3Kα, including but notlimited to breast cancer cells (e.g., MDA-MB-361 and T47D), and cellscomprising one or more mutations in PTEN including but not limited toprostate cancer cells (e.g., PC3), are typically grown in cell growthmedia such as DMEM supplemented with fetal bovine serum and/orantibiotics, and grown to confluency. Cells are then treated withvarious concentrations of test compound for about 2 hours andsubsequently lysed in cell lysis buffer. Lysates are subjected toSDS-PAGE followed by Western blot analysis to detect downstreamsignaling markers, including but not limited to pAKT(S473), pAKT(T308),pS6, and p4E-BP1. Degree of proliferation (and proliferation inhibition)can also be measured for cells at various doses of compound of thepresent invention such as compound 54. Based on percent inhibition ofpAKT and proliferation indicated by these results, IC50 values arecalculated. FIG. 2 provides sample Western blots for downstreamsignaling markers of PI3K activity for a compound of the presentinvention at concentrations of 0.1 μM, 1 μM, and 10 μM, as well ascalculated results for IC50 of pAKT and proliferation inhibition, whichshows inhibition or the PI3K pathway.

Example 39: In Vitro Inhibition of Angiogenesis

Inhibition of angiogenesis in the presence of test compound is evaluatedusing a tube formation assay, such as by using a tube formation assaykit (e.g., commercially available from Invitrogen). Angiogenic capacitycan be measured in vitro using an endothelial cell line, such as humanumbilical vein endothelial cells (HUVEC). The assay is conductedaccording to the kit instructions, in the presence or absence ofcompound. Briefly, a gel matrix is applied to a cell culture surface,cells are added to the matrix-covered surface along with growth factors,with some samples also receiving an inhibitor compound, cells areincubated at 37° C. and 5% CO₂ long enough for control samples (nocompound added) to form tube structures (such as overnight), cells arestained using a cell-permeable dye (e.g., calcein), and cells arevisualized to identify the degree of tube formation. Any decrease intube formation relative to un-inhibited control cells is indicative ofangiogenic inhibition. FIG. 3 shows the results of a sample tubeformation assay, comparing control (no compound added), cells receivinga non-specific PI3K inhibitor (Pan-PI3K Inh), and cells receiving acompound of the invention such as compound 54. Based on doses tested andthe corresponding degree of tube formation inhibition. IC50 values fortube formation are calculated. FIG. 3 also provides IC50 values for cellviability, which can be measured using any number of methods known inthe art, such as staining methods that distinguish live from dead cells(e.g., Image-iT DEAD Green viability stain commercially available fromInvitrogen). FIG. 3 further shows IC50 values for compound 54 andPan-PI3K inhibitor molecules with respect to PI3K α, β, γ, δ, which aredetermined using methods described above, such as in example 5.Generally, the results show that compound 54 has a comparable to higherpotency with respect to angiogenesis inhibition by comparison to thePan-PI3K inhibitor, and is achieved with a compound having specificityfor PI3K α inhibition.

Example 40: In Vivo Efficacy in Xenogeneic Mouse Model of Breast Cancer

Nude mice harboring tumors derived from implantation of human breastadenocarcinoma cells MDA-MB-361 (PI3Kα/HER2 carcinoma) are separatedinto untreated control (vehicle only) and treatment groups. Mice in thetreatment group are further divided into mice receiving 70 mg/kg (70mpk) of a Pan-PI3K inhibitor, or 30 mpk or 60 mpk of test compound. Micein the treatment group receive the defined dose daily by oral lavage for20 to 50 days, during which time tumor weight is calculated as describedin example 37. Blood glucose is monitored periodically followingadministration of treatment. 2 hours after the final treatment, tumorsare harvested and proteins are analyzed by Western blot as describedabove. The effect of the compounds on the localization/viability ofmarginal zone B cells in the spleen is also evaluated at the conclusionof treatment. FIG. 4 shows sample results for tumor size, glucoselevels, and levels of various markers for kinase activity. In general,the results indicate that a compound of the present invention includingcompound 54 has comparable kinase- and tumor growth-inhibition activityto the Pan-PI3K inhibitor, without the impact on blood glucose seen forthe latter. Results of a similar experiment with regard to tumor sizeare reiterated in FIG. 5, followed over a longer period. FIG. 5 furthershows that, in contrast to the Pan-PI3K inhibitor, a compound of thepresent invention including compound 54 does not affectlocalization/viability of marginal zone B cells.

A similar experiment using 786-O cells, a human kidney carcinoma cellline having a non-mutated PI3Kα, instead of MDA cells is used to furtherdemonstrate the specificity of test compounds. For example, a testcompound is compared to a kinase inhibitor with specificity for mTor.The inhibitor with affinity for mTor is effective at inhibiting kinaseactivity (measured by Western blot analysis of downstream markers) andtumor growth in this model, whereas a compound of the present inventionincluding compound 54, while still showing some anti-tumor activity, hasdecreased activity by comparison to that seen in the MDA derived tumors(compare FIG. 4 and FIG. 6). FIG. 6 also provides biochemical potencydata for the two compared inhibitors.

Example 41: Synergistic Combination with Other Kinase Inhibitors

In some embodiments, a compound of the present invention is combinedwith another kinase inhibitor. In some embodiments, the combined kinaseinhibitor is a MEK inhibitor. A median-effect analysis is used todetermine synergism, antagonism, or additivity of a compound of thepresent invention when combined with a MEK inhibitor. The CombinationIndex (CI) is determined using the Chou/Talalay equation. IC50 valuesfor each individual compound is determined in a 72 hr CellTiter-Gloassay. For combination assays, drugs are used at their equipotent ratio(e.g., at the ratio of their IC50's). CalcuSyn software (by Biosoft) isused for dose effect analysis.

To further demonstrate synergy between kinase inhibitors, a cell arrestassay is used to determine the effects of inhibitors alone and incombination on the cycle stage of treated cells. HCT116 cells, a humancolon cancer cell line, is treated with DMSO (carrier), 3 μM of acompound of the present invention including compound 54, 0.3 μM ofPD0325901, or a combination of both 3 μM of compound 54 and 0.3 μM of aMEK inhibitor. Cells are then incubated in the presence of DMSO orinhibitor for 20 hours. The number of cells at each stage in the cellcycle is determined and expressed as a percent of the total, with anincrease in the number of cells arrested at pre-G0/G1 indicatingeffective inhibition of cell cycle progression. FIG. 7 shows sampleresults of such an experiment. While all samples receiving inhibitorshow some increase in cell cycle arrest by comparison to DMSO control,samples receiving both inhibitors show a greater number of arrestedcells than expected by merely adding the individual effects (55%compared to 40%). Thus, these results indicate that the combination of acompound of the present invention including compound 54 with otherinhibitors can be synergistic.

1.-53. (canceled)
 54. A method of providing therapeutic benefit in asubject having a cancer selected from lung cancer and gastric cancer byinhibiting phosphatidyl inositol-3 kinases comprising administering tothe subject a therapeutically effective amount of a compound of thefollowing formula:

or a pharmaceutically acceptable salt thereof, wherein W¹ is CR³; W² isCR⁴; W³ is N; W⁴ is N; W⁵ is CR⁷; W⁶ is CR⁸; R¹ and R² are independentlyhydrogen, alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkoxy,heterocycloalkyloxy, amido, amino, acyl, acyloxy, alkoxycarbonyl,sulfonamido, halo, cyano, hydroxy, nitro, phosphate, urea, or carbonate;R³ is amido of formula —C(O)N(R)₂, or —NHC(O)R, wherein R is hydrogen,alkyl, cylcloalkyl, aryl, heteroaryl, or heteroalicyclic; or whereinsaid (R)₂ groups taken together with the nitrogen to which they areattached form an optionally substituted 4-, 5-, 6-, or 7-membered ring;and R⁴, R⁷ and R⁸ are each hydrogen.
 55. The method of claim 54, whereinR¹ is hydrogen and R² is amino.
 56. The method of claim 55, wherein R²is NH₂.
 57. The method of claim 54, wherein R³ is amido of formula—C(O)N(R)₂ wherein said (R)₂ groups taken together with the nitrogen towhich they are attached form an optionally substituted 4-, 5-, 6-, or7-membered ring.
 58. The method claim 57, wherein R¹ is hydrogen and R²is amino.
 59. The method claim 58, wherein R² is NH₂.
 60. The method ofclaim 59, wherein R³ is amido of formula —C(O)N(R)₂ wherein said (R)₂groups taken together with the nitrogen to which they are attached forman optionally substituted 6-membered ring.
 61. The method of claim 54,wherein the compound or pharmaceutically acceptable salt is


62. The method of claim 54, wherein the cancer is lung cancer.
 63. Themethod of claim 62, wherein the lung cancer is non-small cell lungcancer.
 64. The method of claim 62, wherein the lung cancer is smallcell lung cancer.
 65. The method of claim 54, wherein the cancer isgastric cancer.